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Molecular Biology & Functional Genomics
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4823125001
High Pure FFPE RNA Micro Kit 4823125001
General descriptionLow to medium throughput isolation of total RNA from free FFPE tissue sections in micro scale.ApplicationThe High Pure FFPE RNA Micro Kit isolates total RNA from formalin-fixed, paraffin-embedded (FFPE) tissue samples for direct use in:- Qualitative RT-PCR
- Relative quantification of mRNA with real-time PCR systems such as the LightCycler® 480 System
- Differential display RT-PCR
- cDNA synthesis
- Primer extension
Features and Benefits
The High Pure FFPE RNA Micro Kit isolates total RNA from formalin-fixed, paraffin-embedded (FFPE) tissue samples for direct use in RT-PCR.- Streamline and simplify RNA isolation (even small RNA fragments) from FFPE tissue.
- Obtain a highly concentrated, ready-to-use eluate and excellent recovery of RNA (>80%).
- Isolate DNA-free RNA for use in qualitative and quantitative RT-PCR.
- Minimize RNA loss with a kit that removes contaminants without precipitation or other handling steps that degrade RNA.
- Generate high-quality template RNA that shows excellent performance and linearity in RT-PCR.
Nucleic acids bind to the surface of the glass fiber fleece in the presence of a chaotropic salt (guanidine HCl). This allows the High Pure filter tube to specifically immobilize nucleic acids (both DNA and RNA) while they are freed of contaminants. For RNA isolation, the binding conditions can be optimized to ensure immobilization of all the RNA.
Size Distribution: The typical size of RNA isolated from formalin-fixed tissue ranges from 150 to 1500 bases. However, section thickness, tissue type, age of sample, and the fixation protocol used can affect the yield and quality of the isolated RNA.
Capacity: The High Pure Micro Filter Tubes hold up to 500 µl sample volume.
Sample Material: 1 - 10 µm sections from formalin-fixed, paraffin-embedded (FFPE) tissue (e.g., from colon, breast, liver, kidney, spleen of mammalian species).
Components- Tissue Lysis Buffer
- Proteinase K, recombinant, PCR Grade
- Binding Buffer
- Wash Buffer I
- Wash Buffer II
- DNase I, recombinant, lyophilized
- DNase Incubation Buffer
- Elution Buffer
- High Pure Micro Filter Tubes
- Collection Tubes
Quality
Formalin-fixed, paraffin-embedded tissue sections are homogenized by overnight Proteinase K digestion and purified as described. RNA yield is determined by measuring the optical density at 260 nm.
The RNA eluate and specific primers for the 2M gene are used in one-step RT-PCR. In the following PCR on the LightCycler® 2.0 Instrument (accomplished using the LightCycler® RNA Amplification Kit SYBR Green I and specific primers for 2M), the expected amplification signal is obtained at a Cp-value less than 24.
Absence of contaminating genomic DNA is examined by PCR on a LightCycler® 2.0 Instrument without a reverse transcriptase step; no amplification product is obtained.
Preparation Note
To prepare tissue sections for RNA isolation, fixation reagents must be removed from the samples; after deparaffinization, the sections are ready to be processed with the High Pure FFPE RNA Micro Kit. Deparaffinized tissue samples are disrupted and homogenized during incubation with Proteinase K and a chaotropic salt (guanidine HCl). The homogenate is then applied to the glass fiber fleece in a High Pure Micro Filter Tube.
Under the buffer conditions used in the procedure, all nucleic acids bind specifically to the glass fiber fleece, while contaminating substances (salts, proteins, and other tissue contaminants) do not. DNA in the preparation is digested with DNase I directly on the filter. Brief wash-and-spin steps readily remove the digested DNA fragments and other contaminating substances. The remaining purified RNA is then eluted in a small volume of low-salt buffer.
Analysis Note
Typical RNA Recovery
Starting Material and Quantity: 1 - 10 µm FFPE sections, colon, breast, liver, kidney, spleen of mammalian species
Yield/Recovery: 1.5 - 3.5 µg/5 µm section
Time Required: 60 minutes without 3 hour incubation
Number of Reactions: 50/1-10 µm sectionsOther NotesHigh Pure Technology and Silica Adsorption Kits,
For general laboratory use.Legal InformationLightCycler is a registered trademark of RocheПараметрыQuality Level 100, Торговая марка Roche packaging kit of for 50 isolations
Safety Informationpictograms GHS05,GHS07,GHS08 signalword Danger hcodes H302 + H312 + H332 - H314 - H317 - H334 - H335 - H412 pcodes P260 - P280 - P303 + P361 + P353 - P304 + P340 + P310 - P305 + P351 + P338 + P310 - P342 + P311 Target Organs Respiratory system RIDADR NONH for all modes of transport WGK Germany WGK 2 Flash Point F does not flash Flash Point C does not flash -
5080576001
High Pure miRNA Isolation Kit 5080576001
General descriptionIn the presence of a chaotropic salt, RNA binds selectively to special glass fibers prepacked in the High Pure Spin Filter Tube. Bound RNA is purified in a series of rapid wash-and-spin steps to remove contaminating salts, proteins, and other cellular impurities, and then eluted using a low-salt solution. By lowering the concentration of Binding Enhancer during the binding step in the two-column protocol, the small RNA-containing miRNA passes the first column unbound. When the concentration of Binding Enhancer is increased, the small RNA fraction can be bound to a second High Pure Spin Filter Tube.
Low to medium throughput miRNA isolation.
The High Pure miRNA Isolation Kit purifies and enriches small RNAs, such as microRNA (miRNA) from animal cells and tissue samples (including formalin-fixed, paraffin-embedded sections) or plant material. It can also be used to purify total RNA or to prepare samples enriched for small RNAs (<100 nucleotides).
MicroRNAs are natural regulators of gene expression, affecting almost every cellular process (growth and development, cell differentiation, cell death). They occur naturally in diverse animal and plant species, and hundreds have been discovered since the late 1990s. A single miRNA can ramp down expression of multiple genes. Nucleic acids bind to the surface of the glass fiber fleece in the presence of a chaotropic salt (guanidine HCl). This allows the High Pure filter tube to specifically immobilize nucleic acids (both DNA and RNA) while they are freed of contaminants.
Capacity: The High Pure Spin Filter Tubes hold up to 700 µL sample volume.ApplicationThe High Pure miRNA Isolation kit rapidly purifies RNA that is suitable for direct use in many downstream applications:- Northern blotting
- cDNA synthesis
- Primer extension
- miRNA array hybridization
- Relative quantification of miRNA with RT-PCR using, for example, the LightCycler® 480 System
Features and Benefits- Isolate DNA-free miRNA ideal for qualitative and quantitative reverse transcription. ideal for qualitative and quantitative reverse transcription.
- Obtain excellent performance and linearity in RT-PCR.
- Generate stable , highly pure miRNAs using a simple, efficient protocol.
- Purify miRNA without using hazardous organic solvents. Avoid phenol/chloroform steps required by other suppliers miRNA purification kits.
- Choose one flexible kit for all your miRNA purifications. Use the same kit to purify small RNAs from a variety of sample types.
Components- Tissue Lysis Buffer (for FFPE sections)
- Proteinase K, recombinant (for FFPE sections)
- Binding Buffer
- Binding Enhancer
- Wash Buffer
- Elution Buffer
- High Pure Filter Tubes
- Collection Tubes
Quality
More than 60% recovery is obtained when 1 µg miRNA is added to106 K-562 cells and isolated with the one-column method, and more than 50% is recovered with the two-column method.Other NotesHigh Pure Technology and Silica Adsorption Kits,
For life science research only. Not for use in diagnostic procedures.Legal InformationLightCycler is a registered trademark of RocheПараметрыQuality Level 100, Торговая марка Roche packaging kit of for 50 isolations
Safety Informationpictograms GHS05,GHS07,GHS08 signalword Danger hcodes H302 + H312 + H332 - H314 - H317 - H334 - H335 - H360Df - H412 pcodes P201 - P260 - P280 - P303 + P361 + P353 - P304 + P340 + P310 - P305 + P351 + P338 + P310 - P308 + P313 Target Organs Respiratory system RIDADR NONH for all modes of transport WGK Germany WGK 2 Flash Point F 285.8 °F Flash Point C 141 °C -
High Pure PCR Cleanup Micro Kit
Кат. номер 4983955001 4983912001 General descriptionNucleic acids bind to the surface of the glass fiber fleece in the presence of a chaotropic salt (guanidine HCl). This allows the High Pure filter tube to specifically immobilize nucleic acids (both DNA and RNA) while they are freed of contaminants. The amount of DNA recovered is dependent on the amount of DNA applied to the glass fiber fleece, the elution volume, and the length of the amplification/DNA products. When 5 - 25 µg DNA is applied to the kits High Pure Micro Filter Tube, approximately 85% of the DNA can be recovered.
Capacity of High Pure Micro Filter Tubes: The High Pure Micro Filter Tubes hold up to 500 µL sample volume.
The High Pure PCR Cleanup Micro Kit efficiently purifies products from PCR and other reactions. The kit eliminates primers, mineral oil, salts, unincorporated nucleotides, and thermostable DNA polymerases, which may inhibit subsequent enzymatic reactions such as labeling, sequencing, or cloning of PCR products.ApplicationThe High Pure PCR Cleanup Micro Kit is used to isolate PCR products from amplification and other reactions, and can be used in many molecular biology applications:- Restriction enzyme digests
- Alkaline phosphatase treatment
- Kinase reactions
- Nonradioactive labeling
The kit can also be used to:- Purify cDNA
- Concentrate dilute nucleic acid solutions
- Recover DNA from agarose gel slices
Use one kit for a variety of applications.
The fast and simple High Pure protocols use a tabletop centrifuge to bind, wash, and elute the reaction product down to 10 µL (micro format) in as little as 10 minutes. The procedure conveniently eliminates a concentration step, and is ideal for downstream applications such as labeling, sequencing, cloning, ligation, or amplification using PCR. The purified DNA can also be used for Southern blotting and in vitro transcription.
Features and Benefits- Conserve resources and save time
- Obtain purified product in a small elution volume
- Generate contaminant-free DNA
- Selectively isolate specific DNA fragment sizes
- Eliminate the use of hazardous organic compounds
Components- Binding Buffer
- Binding Enhancer
- Wash Buffer Concentrate
- Elution Buffer
- High Pure Micro Filter Tubes
- Collection Tubes
Quality
Greater than 70% recovery is obtained when 3 µg Roche DNA Molecular Weight Marker VI is applied to the kits High Pure Micro Filter Tubes. Gel electrophoresis of the eluate after purification confirms the complete removal of primer-dimers. The eluted, purified DNA shows no inhibition of amplification using a LightCycler® Instrument with the LightCycler® FastStart DNA MasterPLUS SYBR Green I.
Preparation Note
Nucleic acids bind specifically to the surface of glass fibers in the presence of chaotropic salts. Since the binding process is specific for nucleic acid, the bound material can be separated and purified from impurities by a simple wash step. The Binding Enhancer enables the modification of DNA fragment size exclusions. Small oligonucleotides and dimerized primers from amplification reactions are selectively removed. The nucleic acids elute from the glass fiber fleece in a low-salt buffer or water.
Analysis Note
A 341 bp PCR fragment of the tPA gene was amplified according to a standard block cycler protocol. The resulting reaction mixes were pooled and purified with the High Pure PCR Cleanup Micro Kit. Different amounts of Binding Enhancer were used in the purification procedure. Portions of the PCR product (250 ng each, lanes 2 – 5) and the PCR product mix (16 µL each, lanes 6 – 7) were analyzed on a 1% agarose gel (see Figure 1).
The yield from each purification is shown in Table 1.Other NotesHigh Pure Technology and Silica Adsorption Kits,
For life science research only. Not for use in diagnostic procedures.Legal InformationLightCycler is a registered trademark of RocheПараметрыQuality Level 100, Торговая марка Roche packaging pkg of 200 purifications (04983912001), pkg of 50 purifications (04983955001)
Safety Informationpictograms GHS05,GHS07,GHS08 signalword Danger hcodes H302 + H332 - H314 - H360Df - H412 pcodes P201 - P280 - P303 + P361 + P353 - P304 + P340 + P310 - P305 + P351 + P338 + P310 - P308 + P313 RIDADR NONH for all modes of transport WGK Germany WGK 2 Flash Point F 285.8 °F Flash Point C 141 °C -
High Pure PCR Product Purification Kit
Кат. номер 11732676001 11732668001 General descriptionNucleic acids bind to the surface of the glass fiber fleece in the presence of a chaotropic salt (guanidine HCl). This allows the High Pure filter tube to specifically immobilize nucleic acids (both DNA and RNA) while they are freed of contaminants.
Capacity: The High Pure SpinFilter Tubes hold up to 700 µl sample volume.
Volume: 100 µl
Typical samples include:- Products from amplification or cDNA synthesis reactions
- Enzymatically treated DNA
- DNA from agarose slices
- Dilute nucleic acid solutions
- RNA from transcription reactions
The High Pure PCR Product Purification Kit efficiently and conveniently isolates PCR products from amplification reactions and purifies nucleic acids from other modification reactions. It is also recommended for the purification of cDNA.ApplicationThe High Pure PCR Product Purification Kit is designed for the preparation of concentrated, purified DNA, and can be used directly for most molecular biology applications:- Labeling
- Sequencing
- Cloning
- Restriction enzyme digests
- Alkaline phosphatase treatment
- Kinase reactions
The kit eliminates primers, mineral oil, salts, unincorporated nucleotides, and thermostable DNA polymerases, which may inhibit subsequent enzymatic reactions. It can also be applied to concentrate dilute nucleic-acid solutions. Use one kit for a variety of applications.
The fast and simple High Pure protocols use a tabletop centrifuge to bind, wash, and elute the reaction product down to 10 µl (micro format) in as little as 10 minutes. The procedure conveniently eliminates a concentration step, and is ideal for downstream applications such as labeling, sequencing, cloning, ligation, or amplification using PCR.
Features and Benefits- Quickly purifies multiple PCR products
- Efficiently recover DNA fragments
- Minimize DNA loss
- Eliminate the use of hazardous organic compounds
Components- Binding Buffer
- Wash Buffer
- Elution Buffer
- High Pure Spin Filter Tubes (containing glass fiber fleece)
- Collection Tubes
Quality
More than 70% recovery is obtained when 10 μg DNA Molecular Weight Marker VIII mixed with 16 μg bovine serum albumin are applied to the High Pure Filter Tubes. Gel electrophoresis of the DNA eluate confirms the complete removal of protein and DNA fragments smaller than 100 bp.
Preparation Note
The sample is mixed with a chaotropic salt and applied to the glass fiber fleece in a High Pure Spin Filter Tube. Under the buffer conditions used in the procedure, all nucleic acids (NA) in the sample bind to the glass fleece in the High Pure tube, while contaminating substances (salts, proteins, nucleotides, mineral oil and other contaminants) do not. Brief wash-and-spin steps readily remove these contaminants. Once purified, the NA can be easily eluted in a small volume of low salt buffer.Other NotesHigh Pure Technology and Silica Adsorption Kits,
For life science research only. Not for use in diagnostic procedures.ПараметрыQuality Level 100, Торговая марка Roche packaging pkg of 250 purifications (11732676001), pkg of 50 purifications (11732668001)
Safety Informationpictograms GHS05,GHS07 signalword Danger hcodes H302 + H332 - H314 - H412 pcodes P261 - P273 - P280 - P303 + P361 + P353 - P304 + P340 + P310 - P305 + P351 + P338 + P310 RIDADR NONH for all modes of transport WGK Germany WGK 2 -
11796828001
High Pure PCR Template Preparation Kit 11796828001
General descriptionLow to medium throughput genomic DNA isolation.
High Pure PCR Template Preparation Kit; Instructions For Use,
Nucleic acids bind to the surface of the glass fiber fleece in the presence of a chaotropic salt (guanidine HCl). This allows the High Pure filter tube to specifically immobilize nucleic acids (both DNA and RNA) while they are freed of contaminants.
Note: A special Inhibitor Removal Buffer is included which allows the use of heparinized sample material (100 U/mL of heparin). This buffer increases the sensitivity and reproducibility of assays performed with the isolated nucleic acid, even when the sample contains heparin.
Capacity: The High Pure Spin Filter Tubes hold up to 700 µL sample volume.ApplicationHigh Pure PCR Template Preparation Kit has been used in the extraction of genomic DNA from whole blood samples, cervical lesion specimens, gastric muscosal tissue and cervical carcinoma cell lines.
The High Pure PCR Template Preparation Kit purifies nucleic acids from a wide variety of sample materials including whole blood, cultured cells, and tissue. The isolated nucleic acids can be used for:- Long-template PCR
- Real-time, quantitative PCR
- SNP detection
- Southern blotting
- Cloning
Features and Benefits
The High Pure PCR Template Preparation Kit purifies nucleic acids from a wide variety of sample materials, including whole blood, cultured cells, and tissue samples. Bacteria and yeast require a specific pre-lysis treatment with lysozyme or lyticase.- Minimize DNA loss using a kit that removes contaminants without precipitation or other handling steps that degrade DNA.
- Recover high molecular weight DNA (30 to 50 kb) that is suitable for long-template PCR.
- Improve reliability and reproducibility in downstream applications (real-time, quantitative PCR).
- Save time and maximize flexibility by preparing multiple PCR templates simultaneously.
- Eliminate the use of hazardous organic compounds such as cesium chloride, phenol, chloroform, and ethidium bromide.
Components- Tissue Lysis Buffer
- Binding Buffer
- Proteinase K, recombinant PCR grade
- Inhibitor Removal Buffer
- Washing Buffer
- Elution Buffer
- High Pure Filter Tubes
- Collection Tubes
Quality
DNA is isolated from 25 mg of calf thymus, 1 x 106 K562 cells, and 200 µl of EDTA whole blood. Yield is measured via OD for tissue and cell samples. The quality of the nucleic acid is controlled in an Expand Long Range PCR with a 9.3 kb amplification product for DNA derived from cells. PCR on a LightCycler® Instrument is performed on human blood research samples using kits for Factor V Leiden and CycA.
Principle
Blood cells or tissue are lysed by a short incubation with a special Lysis Buffer and Proteinase K in the presence of a chaotropic salt such as guanidine-HCl, which immediately inactivates all nucleases. Cellular nucleic acids bind selectively to special glass fibers pre-packed in the High Pure Purification Filter Tube. Bound nucleic acid is purified in a series of rapid wash-and-spin steps to remove contaminating cellular components. Finally, low salt elution releases the nucleic acid from the glass fiber. This simple method eliminates the need for organic solvent extractions and DNA precipitation, allowing for rapid purification of many samples simultaneously.Legal InformationLightCycler is a registered trademark of RocheПараметрыQuality Level 100, Торговая марка Roche packaging pkg of 100 purifications
Safety Informationpictograms GHS05,GHS07,GHS08 signalword Danger hcodes H302 + H332 - H315 - H317 - H318 - H334 - H335 - H412 pcodes P261 - P264 - P280 - P304 + P340 + P312 - P305 + P351 + P338 + P310 - P342 + P311 Target Organs Respiratory system RIDADR NONH for all modes of transport WGK Germany WGK 2 Flash Point F does not flash Flash Point C does not flash -
High Pure Plasmid Isolation Kit
Кат. номер 11754785001 11754777001 General descriptionLow to medium throughput, mini scale, plasmid isolation.
The High Pure Plasmid Isolation Kit isolates purified plasmid DNA in small quantities using the alkaline lysis method, a commonly used method that generates highly purified plasmid DNA from E. coli, free of RNA contamination.
Nucleic acids bind to the surface of the glass fiber fleece in the presence of a chaotropic salt (guanidine HCl). This allows the High Pure filter tube to specifically immobilize nucleic acids (both DNA and RNA) while they are freed of contaminants.
Capacity: The High Pure Spin Filter Tubes hold up to 700 µL sample volume.
Sample: 0.5 - 4.0 mL cultures of E. coli (harvested at a density of 1.5 - 5.0 A 600 units/mL)ApplicationThe High Pure Plasmid Isolation Kit prepares up to 15 µg purified plasmid DNA from bacterial cultures, that can be used directly in most molecular biology applications:- PCR
- Sequencing
- Cloning
- In vitro transcription
- Restriction enzyme digests
- Random primed labeling
Features and Benefits- Quickly purify up to 24 plasmid samples in <30 minutes.
- Minimize DNA loss with a kit that removes contaminants without precipitation or other handling steps that degrade DNA.
- Improve reliability and reproducibility in downstream applications with a kit that removes RNA and other impurities that cause plasmid DNA to behave unpredictably.
- Eliminate the use of hazardous organic compounds such as cesium chloride, phenol, chloroform, and ethidium bromide.
Components- Suspension Buffer
- RNase A, dry powder
- Lysis Buffer
- Binding Buffer
- Wash Buffer I
- Wash Buffer II
- Elution Buffer
- High Pure Spin Filter Tubes (containing glass fiber fleece)
- Collection Tubes
Quality
Plasmid pUC19 (4.5 µg) is purified from a 1.5 mL suspension of E. coli JM83, which was grown in LB-medium with ampicillin for 16 hours, to a cell density of 5 A 600 units/mL. 1 µg of the purified plasmid DNA is incubated for 1 hour at +37°C with 5 units of the restriction endonuclease Eco RI and then analyzed by agarose gel electrophoresis. The isolated plasmid DNA is as sensitive to restriction endonuclease digestion as plasmid DNA isolated by CsCl density centrifugation.
Preparation Note
The kit relies on alkaline lysis to release plasmid DNA from bacteria. RNase removes all RNA in the lysate. After cellular debris and (entrapped) genomic DNA are removed by centrifugation, the remaining supernatant is mixed with a chaotropic salt and applied to the glass fiber fleece in a High Pure Spin Filter Tube. Under the buffer conditions used in the procedure, the plasmid binds to the glass fiber fleece, while contaminating substances (salts, proteins, and other cellular contaminants) do not. Brief wash-and-spin steps readily remove these contaminants. Once purified, the plasmid can be easily eluted in a small volume of low-salt buffer or water.Other NotesHigh Pure Technology and Silica Adsorption Kits,
For life science research only. Not for use in diagnostic procedures.ПараметрыQuality Level 100, Торговая марка Roche packaging pkg of 250 purifications (11754785001), pkg of 50 purifications (11754777001)
Safety Informationpictograms GHS05,GHS07 signalword Danger hcodes H302 + H332 - H314 pcodes P261 - P280 - P301 + P330 + P331 - P303 + P361 + P353 - P304 + P340 + P310 - P305 + P351 + P338 + P310 RIDADR UN 1824 8 / PGIII WGK Germany WGK 1 Flash Point F Not applicable Flash Point C Not applicable -
10223565001
Histone 10223565001
General descriptionMixture of histones H1, H2A, H2B, H3, and H4, isolated from calf thymus.
Histones are a group of DNA-binding proteins that are characterized by relatively high levels of lysine and arginine. Five different fractions of histones have been isolated and characterized. These are named as H1, H2A, H2B, H3 and H4. The H1 fraction is lysine-rich, while the H2A and H2B are slightly lysine-rich, The H3 and H4 fractions are arginine-rich. The molecular weights of histones are approximately 11 to 21 kDa depending on the fraction.ApplicationHistone from calf thymus has been used:- in in vitro kinase assay to check the phosphorylation of histones by protein kinase A.
- to check the in vitro methyltransferase activity of protein argininemethyltransferases from Oryza sativa (OsPRMTs).
- in anti-histone ELISA.
Biochem/physiol Actions
Histones have an important role in the organization and modification of chromatin. The nucleosome is the basic unit of the chromatin. It is made up of two molecules each of histones H2A, H2B, H3 and H4, wrapped with approximately 146bp of DNA. Histone H1 binds to the nucleosomes at the DNA entry and exit points and stabilizes them.
Physical form
Lyophilizate (natural mixture of histones H1, H2A, H2B, H3, and H4).
Preparation Note
Working concentration: 1 to 5 mg/ml.
Working solution: Recommended solvent is double-distilled water.
Storage conditions (working solution): Aqueous solution is stable at -15 to -25 °C for several months.
Avoid repeated freezing and thawing!Other NotesFor life science research only. Not for use in diagnostic procedures.ПараметрыQuality Level 100, form lyophilized packaging pkg of 10 mg Торговая марка Roche shipped in wet ice storage temp. 2-8°C
Safety InformationRIDADR NONH for all modes of transport WGK Germany WGK 1 Flash Point F Not applicable Flash Point C Not applicable -
11034758001
Histone H3 11034758001
General descriptionLyophilizate.
Mammals contain three main classes of histone H3 variants: the replicative histones (H3.1 and H3.2), the replacement histone (H3.3) and the centromeric histone (Cenp-A). Histone H3 is located on human chromosome 6p22. H3.1 is also called as HIST1H3E. It is mainly expressed in the S phase.ApplicationHistone H3 has been used to determine the role of histones in metastasis. It has also been used in dot blot and western blotting.
Biochem/physiol Actions
Histone proteins are basic building blocks of chromatin. Mutations in Histone H3 results in adult cerebellar high-grade gliomas. It controls protein-protein interactions to induce binding of trans-acting factors that “drive” chromatin condensation. H3.1 acts as a replication-dependent histone.
Quality
Typical analysis: The preparation is electrophoretically homogeneous.
Preparation Note
Working solution: Solvent is recommended in double-distilled water.Other NotesFor life science research only. Not for use in diagnostic procedures.ПараметрыQuality Level 100, form lyophilized packaging pkg of 1 mg Торговая марка Roche shipped in wet ice storage temp. 2-8°C
Safety InformationRIDADR NONH for all modes of transport WGK Germany WGK 1 Flash Point F Not applicable Flash Point C Not applicable -
11691112001
Human Genomic DNA 11691112001
General descriptionDNA is composed of units called nucleotides arranged into two long polynucleotide chains, resulting in a double helical structure. Nucleotides contain a phosphate group, a sugar group and a nitrogen base. There are four types of nitrogen bases namely adenine (A), thymine (T), guanine (G) and cytosine (C). The arrangement of these bases establishes the genetic codon. The difference in the nucleotide sequences is the reason why organisms differ from one another. In eukaryotic cell, the DNA is localized to the nucleus.
High molecular weight (>50kb) genomic DNA isolated from human blood (buffy coat) by the method of Sambrook et al..ApplicationHuman Genomic DNA is suitable for:- Southern hybridization analysis
- genomic library construction
- the amplification of large DNA targets by the Expand System
- to assess the quality or integrity of DNA sample using qPCR or real time (RT) PCR and as a control during DNA sequencing
Biochem/physiol Actions
DNA is an essential component of the mechanism for heredity. The nucleotide sequences carry information regarding different biological processes. The genetic codons encode proteins essential for biological function. This genetic information is transmitted to the next generation during cell division. The amplification of very long fragments of genomic DNA requires template DNA of very high quality. In some cases, amplification failure may be due to poor template quality. This particular quality of genomic DNA is prepared to ensure reliable amplification of long DNA fragments.
Quality
Molecular weight: The preparation is electrophoretically separated on a 0.5% agarose gel and the gel is stained with ethidium bromide. The molecular weight of the purified genomic DNA is greater than 50kb.
Function test: The preparation is used as template in a PCR with the Expand PCR System and appropriate primers from the human tPA Control Primer Set. Amplification products up to 27kb long are obtained.
Absence of contaminating organisms: The serum used for this preparation was tested for HBs antigen and the presence of antibodies to HIV-1, HIV-2, HCV. All tests were negative.
Physical form
Solution in 10 mM Tris HCl, 1 mM EDTA, pH 8.0Other NotesFor life science research only. Not for use in diagnostic procedures.ПараметрыQuality Level 100, form solution packaging pkg of 100 μg Торговая марка Roche concentration 0.2 mg/mL shipped in wet ice storage temp. 2-8°C
Safety InformationRIDADR NONH for all modes of transport Flash Point F No data available Flash Point C No data available -
11363735001
Hybridoma Fusion and Cloning Supplement 11363735001
General descriptionMedium supplement composed of albumin, insulin, transferrin, cytokines, a cholesterol source, other defined organic and inorganic compounds and fetal calf serum (final concentration: 0.5%).
HFCS contains human proteins. The raw material from which the human proteins were isolated, has been tested for the presence of Hepatitis B surface Antigen (HBsAg) and HIV-1/2 antibodies and found to be negative (according to current quality control procedures).ApplicationHybridoma Fusion and Cloning Supplement (HFCS) is used as a supplement to the normal culture medium to support the growth of B-cell hybridomas after fusion and during cloning. HFCS replaces feeder cells. It is also used to optimize the growth of hybridomas after the thawing of cells stored in liquid nitrogen.
Physical form
Solution (50x concentrated, pH 7.4); filtered through 0.2 μm pore size membrane
Preparation Note
Working concentration: HFCS (50x) is diluted 1:50 (v/v) with basal medium. It is strongly suggested that you use RPMI 1640. The final medium should also contain L-glutamine and sodium bicarbonate.
Analysis Note
Biological activity: Assayed for high cloning efficiency of a hybridoma cell line.Other NotesFor life science research only. Not for use in diagnostic procedures.ПараметрыQuality Level 100, sterility 0.2 μm filtered form solution packaging pkg of 10 mL (50x) Торговая марка Roche application(s) cell culture | hybridoma: suitable impurities Mycoplasma, tested, 10 EU/mL Endotoxin (LAL) solubility water: miscible shipped in dry ice storage temp. −20°C
Safety InformationRIDADR NONH for all modes of transport WGK Germany WGK 1 Flash Point F does not flash Flash Point C does not flash -
10843555001
Hygromycin B 10843555001
Empirical Formula (Hill Notation): C20H37N3O13 CAS Number: 31282-04-9 Molecular Weight: 527.52 MDL number: MFCD06795479 PubChem Substance ID: 329749191 General description- Formula: C20H37N3O13
- LD50: 6 mg/kg in mouse (i.v.);13 mg/kg in guinea pig (i.p.); 63 mg/kg in rat (i.p.)
- Molecular weight: Mr = 527.5
- Working concentration: 50 - 1,000 µg/ml in cell culture. A commonly used concentration for the selection of mammalian cells is 200 µg/ml. However, the optimal concentration must be experimentally determined, since it may vary depending on the type of cell used.
ApplicationHygromycin B is an aminoglycosidic antibiotic that inhibits protein synthesis in prokaryotes and eukaryotes. It is used to select and maintain the phenotype of eukaryotic cells that are stably transfected with the E. coli hygromycin-resistance gene (hyg or hph).
Biochem/physiol Actions
Mode of Action: The product acts by inhibiting protein synthesis by inducing the misreading of the m-RNA template in the prokaryote, with the potency to inhibit translation.
Antimicrobial Spectrum: Hygromycin B acts against bacteria, fungi and higher eukaryotic cells.
Quality
Purity: >80% (HPLC)
Unit Definition
Unit Assay: Unit/mg
We do not have this information.
Physical form
Solution, 50 mg/ml in PBS (phosphate-buffered saline), filtered through 0.2 μm pore size membrane
Preparation Note
Working concentration: Recommended concentration for the selection of resistant cells is 50 to 1000 µg/ml in cell culture. A commonly used concentration for the selection of mammalian cells is 200 µg/ml. However, the optimal concentration must be determined experimentally, and may vary with the type of cell used.
Storage and Stability
Store at 2 to 8 °C. (solution)Other NotesFor life science research only. Not for use in diagnostic procedures.
Solution, 50 mg/ml, in PBS (phosphate-buffered saline), filtered through 0.2 μm pore size membrane.Параметрыsterility non-sterile; 0.2 µm filtered assay 80% (HPLC) form buffered aqueous solution composition Hygromycin B, >80% HPLC packaging pkg of 20 mL (1 g) Торговая марка Roche concentration 50 mg/mL application(s) transfection: suitable shipped in wet ice SMILES string CN[C@H]1C[C@@H](N)[C@H](O)[C@@H](O[C@@H]2O[C@H](CO)[C@H](O)[C@@H]3O[C@]4(O[C@H]([C@H](N)CO)[C@H](O)[C@H](O)[C@H]4O)O[C@H]23)[C@@H]1O InChI 1S/C20H37N3O13/c1-23-7-2-5(21)9(26)15(10(7)27)33-19-17-16(11(28)8(4-25)32-19)35-20(36-17)18(31)13(30)12(29)14(34-20)6(22)3-24/h5-19,23-31H,2-4,21-22H2,1H3/t5-,6-,7+,8-,9+,10-,11+,12-,13+,14-,15-,16+,17+,18-,19+,20+/m1/s1 InChI key GRRNUXAQVGOGFE-XKIAHZFYSA-N
Safety Informationpictograms GHS05,GHS06,GHS08 signalword Danger hcodes H301 - H312 - H317 - H318 - H330 - H334 pcodes P260 - P280 - P301 + P310 + P330 - P304 + P340 + P310 - P305 + P351 + P338 + P310 - P342 + P311 - P403 + P233 RIDADR UN 2810 6.1 / PGIII WGK Germany WGK 1 Flash Point F does not flash Flash Point C does not flash -
11719394001
Immunoprecipitation Kit (Protein A) 11719394001
General descriptionKit contains all reagents necessary for cell lysis, solubilization, stabilization, and immunopurification of proteins.
Immunoprecipitation is a widely used method for the analysis of target antigens in complex mixture of proteins. The protein of interest can be concentrated and immunoaffinity -purified in one step on an analytical scale via a specific antibody. Often, immunoprecipitated proteins are functionally fully active and can be further analyzed with respect to enzymatic activity, interactions, modifications and structure.ApplicationImmunoprecipitation Kit (Protein A) has been used for the immunoprecipitation of proteins from cellular extracts with Protein A Agarose.
Packaging
1 kit containing 5 components.
Preparation Note
Working solution: Lysis buffer/wash buffer 1
The kit contains reagents for 125 ml of lysis buffer/wash buffer 1. Prepare at least a minimal volume of 25 ml, sufficient for four immunoprecipitations.
To prepare 25 ml of lysis buffer/wash buffer 1 mix 5 ml core buffer, 3.75 ml NaCl, 2.5 ml detergent mix and 1 cOmplete tablet. Add water to a final volume of 25 ml.
Solution is stable at 2 to 8 °C for 24 hours. When stored in aliquots at -15 to -25 °C, the solution is stable for at least four weeks. Mix thoroughly after thawing.
Wash buffer 2
The kit contains reagents for 50 ml of wash buffer 2. 2 ml of this buffer is required for one immunoprecipitation.
To prepare 50 ml of wash buffer 2 mix 10 ml core buffer, 25 ml NaCl and 0.5 ml detergent mix. Add water to a final volume of 50 ml.
Solution is stable at 2 to 8 °C. For longer periods, store aliquots at -15 to -25 °C. Mix thoroughly after thawing.
Wash buffer 3
The kit contains reagents for 25 ml of wash buffer 3. 1 ml of this buffer is required for one immunoprecipitation.
To prepare 25 ml mix 1 ml core buffer and 0.25 ml detergent mix. Add water to a final volume of 25 ml.
Solution is stable at 2 to 8 °C. For longer periods, store aliquots at -15 to -25 °C. Mix thoroughly after thawing.Other NotesFor life science research only. Not for use in diagnostic procedures.Параметрыusage sufficient for 20 reactions Quality Level 100, Торговая марка Roche shipped in wet ice storage temp. 2-8°C
Safety Informationpictograms GHS05,GHS07,GHS09 signalword Danger hcodes H314 - H317 - H411 pcodes P273 - P280 - P303 + P361 + P353 - P304 + P340 + P310 - P305 + P351 + P338 + P310 - P391 RIDADR NONH for all modes of transport WGK Germany WGK 3 Flash Point F does not flash Flash Point C does not flash -
11719386001
Immunoprecipitation Kit (Protein G) 11719386001
General descriptionKit contains all reagents necessary for cell lysis, solubilization, stabilization, and immunopurification of proteins.
Specificity
Protein G is a cell wall protein, isolated from a specific bacterial strain, which has specific binding sites for certain classes of immunoglobulins from different species. Protein G binds nearly all subclasses of IgG, but no other classes of immunoglobulins.ApplicationThe immunoprecipitation kit has been used for the immunoprecipitation of proteins from cellular extracts with Protein G Agarose.
Packaging
1 kit containing 5 components.
Preparation Note
Working solution: Lysis buffer/wash buffer 1
The kit contains reagents for 125 ml of lysis buffer/wash buffer 1. Prepare at least a minimal volume of 25 ml, sufficient for four immunoprecipitations.
To prepare 25 ml of lysis buffer/wash buffer 1 mix 5 ml core buffer, 3.75 ml NaCl, 2.5 ml detergent mix and 1 cOmplete Tablet. Add water to a final volume of 25 ml.
Solution is stable at 2 to 8 °C for 24 hours. When stored in aliquots at -15 to -25 °C, the solution is stable for at least four weeks. Mix thoroughly after thawing.
Wash buffer 2
The kit contains reagents for 50 ml of wash buffer 2. 2 ml of this buffer is required for one immunoprecipitation.
To prepare 50 ml of wash buffer 2 mix 10 ml core buffer, 25 ml NaCl and 0.5 ml detergent mix. Add water to a final volume of 50 ml.
Solution is stable at 2 to 8 °C. For longer periods, store aliquots at -15 to -25 °C. Mix thoroughly after thawing.
Wash buffer 3
The kit contains reagents for 25 ml of wash buffer 3. 1 ml of this buffer is required for one immunoprecipitation.
To prepare 25 ml mix 1 ml core buffer and 0.25 ml detergent mix. Add water to a final volume of 25 ml.
Solution is stable at 2 to 8 °C. For longer periods, store aliquots at -15 to -25 °C. Mix thoroughly after thawing.Other NotesFor life science research only. Not for use in diagnostic procedures.Параметрыusage sufficient for 20 reactions Quality Level 100, Торговая марка Roche shipped in wet ice storage temp. 2-8°C
Safety Informationpictograms GHS05,GHS07,GHS09 signalword Danger hcodes H314 - H317 - H411 pcodes P273 - P280 - P303 + P361 + P353 - P304 + P340 + P310 - P305 + P351 + P338 + P310 - P391 RIDADR NONH for all modes of transport WGK Germany WGK 3 Flash Point F does not flash Flash Point C does not flash -
11684809910
In Situ Cell Death Detection Kit, AP 11684809910
General descriptionKit for the detection and quantification of apoptotic cell death on a single-cell level by light microscopy in immunohistocytochemistry.
Contents:- Enzyme Solution (TdT)
- Label Solution (fluorescein-dUTP)
- Converter AP (anti-fluorescein antibody-AP), ready-to-use
Specificity
The TUNEL reaction preferentially labels DNA strand breaks generated during apoptosis. This allows discrimination of apoptosis from necrosis and from primary DNA strand breaks induced by cytostatic drugs or irradiation.ApplicationThe In Situ Cell Death Detection Kit, AP, is a precise, fast, and simple nonradioactive technique to detect and quantify apoptotic cell death at the single-cell level in cells and tissues by light microscopy; it is not intended for diagnostic procedures. Thus, the In Situ Cell Death Detection Kit can be used in many different assay systems. Examples are:- Detection of individual apoptotic cells in frozen and formalin-fixed tissue sections in basic research†
- Determination of sensitivity of malignant cells to drug-induced apoptosis in cancer research
- Typing of cells undergoing cell death in heterogeneous populations by double staining procedures
Features and Benefits- Sensitive: The maximum intensity of labeling (cell staining) of apoptotic cells is higher than the nick translation method
- Fast: The use of fluorescein-dUTP allows analysis of the samples directly after the TUNEL reaction, but before the addition of the secondary detection system
- Convenient: The direct labeling procedure using fluorescein-dUTP allows verification of the efficiency of the TUNEL reaction during the assay procedure
- Accurate: Identification of apoptosis at a molecular level (DNA-strand breaks) and identification of cells at the very early stages of apoptosis
- Flexible: No substrate included; provides the opportunity to select the staining procedure of choice
Packaging
1 kit containing 3 components.
Specifications
Widely used methods to determine apoptosis include the analysis of the genomic DNA by agarose-gel electrophoresis and DNA fragmentation assays based on 3H-thymidine and, alternatively, 5-Bromo-2-deoxy-uridine. The methods involve the separation of fragmented, low molecular-weight DNA from unfragmented, high molecular-weight DNA in a given cell population. Thus, these methods do not provide information about the fate of an individual cell in a given cell population or, particularly, in tissue sections. Alternatively, individual apoptotic cells may be microscopically recognized because of the characteristic appearance of nuclear chromatin condensation and fragmentation, but this method is subjective and limited to a relatively narrow time window when the morphological changes are at a maximum.
The hallmark of apoptosis is DNA degradation, which in early stages is selective to the internucleosomal DNA linker regions. The DNA cleavage may yield double-stranded and single-stranded DNA breaks (nicks). Both types of breaks can be detected by labeling the free 3-OH termini with modified nucleotides (e.g., biotin-dUTP, DIG-dUTP, fluorescein-dUTP) in an enzymatic reaction. The enzyme terminal deoxynucleotidyl transferase (TdT) catalyzes the template-independent polymerization of deoxyribonucleotides to the 3-end of single- and double-stranded DNA. This method has also been termed TUNEL (TdT-mediated dUTP-X nick end labeling). Alternatively, free 3-OH groups may be labeled using DNA polymerases by the template-dependent mechanism called nick translation. However, the TUNEL method is considered to be more sensitive and faster.
Sample material: Cytospin and cell smear preparations, adherent cells grown on slides, and frozen and paraffin-embedded tissue sections.
Principle
The In Situ Cell Death Detection Kit, AP is based on the detection of single- and double-stranded DNA breaks that occur at the early stages of apoptosis.
Apoptotic cells are fixed and permeabilized. Subsequently, the cells are incubated with the TUNEL reaction mixture that contains TdT and fluorescein-dUTP. During this incubation period, TdT catalyzes the addition of fluorescein-dUTP at free 3-OH groups in single- and double-stranded DNA. After washing, the label incorporated at the damaged sites of the DNA is marked by an anti-fluorescein antibody conjugated with the reporter enzyme alkaline phosphatase. After washing to remove unbound enzyme conjugate, the AP retained in the immune complex is visualized by a substrate reaction.
Preparation Note
Working solution: Add total volume (50 µl) of Enzyme Solution to the remaining 450 µl Label Solution to obtain 500 µl TUNEL reaction mixture.
Mix well to equilibrate components.
Storage conditions (working solution): TUNEL reaction mixture
The TUNEL reaction mixture should be prepared immediately before use and should not be stored. Keep TUNEL reaction mixture on ice until use.
Converter-AP
Once thawed the Converter-AP solution should be stored at 2 to 8 °C (maximum stability
6 months).
Note: Do not freeze.Other NotesFor life science research only. Not for use in diagnostic procedures.Параметрыusage sufficient for 50 tests Quality Level 100, Торговая марка Roche shipped in dry ice storage temp. 20°C
Safety Informationpictograms GHS07,GHS08,GHS09 signalword Danger hcodes H317 - H350i - H411 pcodes P201 - P261 - P273 - P280 - P308 + P313 - P391 RIDADR UN 1556 6.1 / PGIII WGK Germany WGK 3 Flash Point F does not flash Flash Point C does not flash -
11684795910
In Situ Cell Death Detection Kit, Fluorescein 11684795910
General descriptionKit for the detection and quantification of apoptosis at the single-cell level, based on labeling of DNA strand breaks (TUNEL technology); analysis by fluorescence microscopy or flow cytometry.
Widely used methods to determine apoptosis include the analysis of the genomic DNA by agarose-gel electrophoresis and DNA fragmentation assays based on 3H-thymidine and, alternatively, 5-Bromo-2-deoxy-uridine. The methods involve the separation of fragmented, low molecular weight DNA from unfragmented, high molecular weight DNA in a given cell population. Thus, these methods do not provide information about the fate of an individual cell in a given cell population, or particularly, in tissue sections. Alternatively, individual apoptotic cells may be microscopically recognized because of the characteristic appearance of nuclear chromatin condensation and fragmentation, but this method is subjective and limited to a relatively narrow time window when the morphological changes are at a maximum.
The hallmark of apoptosis is DNA degradation, which in early stages, is selective to the internucleosomal DNA linker regions. The DNA cleavage may yield double-stranded and single-stranded DNA breaks (nicks). Both types of breaks can be detected by labeling the free 3-OH termini with modified nucleotides (e.g., biotin-dUTP, DIG-dUTP, fluorescein-dUTP) in an enzymatic reaction. The enzyme terminal deoxynucleotidyl transferase (TdT) catalyzes the template-independent polymerization of deoxyribonucleotides to the 3-end of single- and double-stranded DNA. This method has also been termed TUNEL (TdT-mediated dUTP-X nick end labeling). Alternatively, free 3-OH groups may be labeled using DNA polymerases by the template-dependent mechanism called nick translation. However, the TUNEL method is considered to be more sensitive and faster.
Specificity
The TUNEL reaction preferentially labels DNA strand breaks generated during apoptosis. This allows discrimination of apoptosis from necrosis and from primary DNA strand breaks induced by cytostatic drugs or irradiation.ApplicationThe In Situ Cell Death Detection Kit, Fluorescein, is a precise, fast, and simple nonradioactive technique to detect and quantify apoptotic cell death at the single-cell level by fluorescence microscopy and quantitative detection by flow cytometry in cells and tissues. Thus, the In Situ Cell Death Detection Kit can be used in many different assay systems.
Examples are:- Detection of individual apoptotic cells in frozen and formalin-fixed tissue sections in basic research
- Determination of sensitivity of malignant cells to drug-induced apoptosis in cancer research
- Typing of cells undergoing cell death in heterogeneous populations by double staining procedures
Features and Benefits- Sensitive: The direct labeling procedure using fluorescein-dUTP reduces background labeling
- Fast: The use of fluorescein-dUTP allows analysis of the samples directly after the TUNEL reaction
- Convenient: No secondary detection system required
- Accurate: Identification of apoptosis at a molecular level (DNA-strand breaks) and identification of cells at the very early stages of apoptosis
Packaging
1 kit containing 2 components.
Preparation Note
Working solution: Add total volume (50 µl) of Enzyme Solution to the remaining 450 µl Label Solution to obtain 500 µl TUNEL reaction mixture.
Mix well to equilibrate components.Other NotesFor life science research only. Not for use in diagnostic procedures.Параметрыusage sufficient for 50 tests Quality Level 100, Торговая марка Roche shipped in dry ice storage temp. 20°C
Safety Informationpictograms GHS08,GHS09 signalword Danger hcodes H350i - H411 pcodes P201 - P273 - P280 - P308 + P313 - P391 - P501 RIDADR UN 1556 6.1 / PGIII WGK Germany WGK 3 Flash Point F does not flash Flash Point C does not flash -
11684817910
In Situ Cell Death Detection Kit, POD 11684817910
General descriptionKit for the detection and quantification of apoptotic cell death on a single-cell level by light microscopy in immunohistocytochemistry.ApplicationThe In Situ Cell Death Detection Kit, POD, is a precise, fast, and simple nonradioactive technique to detect and quantify apoptotic cell death at the single-cell level in cells and tissues. Thus, the In Situ Cell Death Detection Kit can be used in many different assay systems.- Examples are: Detection of individual apoptotic cells in frozen, paraffin-embedded and formalin-fixed tissue sections in basic research and routine pathology
- Determination of sensitivity of malignant cells to drug-induced apoptosis in cancer research and clinical oncology
- Typing of cells undergoing cell death in heterogeneous populations by double staining procedures
Features and Benefits
Sensitive: The maximum intensity of labeling (cell staining) of apoptotic cells is higher than the nick translation method.
Fast: The use of fluorescein-dUTP allows analysis of the samples directly after the TUNEL reaction, but before the addition of the secondary detection system.
Convenient: The direct labeling procedure using fluorescein-dUTP allows verification of the efficiency of the TUNEL reaction during the assay procedure.
Accurate: Identification of apoptosis at a molecular level (DNA-strand breaks) and identification of cells at the very early stages of apoptosis.
Flexible: No substrate included; provides the opportunity to select the staining procedure of choice.
Packaging
1 kit containing 3 components.
Principle
The In Situ Cell Death Detection Kit, POD is based on the detection of single- and double-stranded DNA breaks that occur at the early stages of apoptosis.
Apoptotic cells are fixed and permeabilized. Subsequently, the cells are incubated with the TUNEL reaction mixture that contains TdT and fluorescein-dUTP. During this incubation period, TdT catalyzes the addition of fluorescein-dUTP at free 3-OH groups in single- and double-stranded DNA. After washing, the label incorporated at the damaged sites of the DNA is marked by an anti-fluorescein antibody conjugated with the reporter enzyme peroxidase. After washing to remove unbound enzyme conjugate, the POD retained in the immune complex is visualized by a substrate reaction.
Preparation Note
Storage conditions (working solution): TUNEL reaction mixture
The TUNEL reaction mixture should be prepared immediately before use and should not be stored. Keep TUNEL reaction mixture on ice until use.
Converter-peroxidase
Once thawed the Converter-peroxidase solution should be stored at 2 to 8 °C (maximum stability 6 months).
Note: Do not freeze.
Sample material: Cytospin and cell smear preparations, adherent cells grown on slides, and frozen and paraffin-embedded tissue sections.Other NotesFor life science research only. Not for use in diagnostic procedures.Параметрыusage sufficient for 50 tests Quality Level 100, Торговая марка Roche shipped in dry ice storage temp. 20°C
Safety Informationpictograms GHS07,GHS08,GHS09 signalword Danger hcodes H317 - H350i - H411 pcodes P201 - P261 - P273 - P280 - P308 + P313 - P391 RIDADR UN 1556 6.1 / PGIII WGK Germany WGK 3 Flash Point F does not flash Flash Point C does not flash -
12156792910
In Situ Cell Death Detection Kit, TMR red 12156792910
General descriptionWidely used methods to determine apoptosis include the analysis of the genomic DNA by agarose-gel electrophoresis and DNA fragmentation assays based on 3H-thymidine and, alternatively, 5-Bromo-2-deoxy-uridine. The methods involve the separation of fragmented, low molecular weight DNA from unfragmented, high molecular weight DNA in a given cell population. Thus, these methods do not provide information about the fate of an individual cell in a given cell population, or particularly, in tissue sections. Alternatively, individual apoptotic cells may be microscopically recognized because of the characteristic appearance of nuclear chromatin condensation and fragmentation, but this method is subjective and limited to a relatively narrow time window when the morphological changes are at a maximum.
The hallmark of apoptosis is DNA degradation, which in early stages, is selective to the internucleosomal DNA linker regions. The DNA cleavage may yield double-stranded and single-stranded DNA breaks (nicks). Both types of breaks can be detected by labeling the free 3-OH termini with modified nucleotides (e.g., biotin-dUTP, DIG-dUTP, fluorescein-dUTP) in an enzymatic reaction. The enzyme terminal deoxynucleotidyl transferase (TdT) catalyzes the template-independent polymerization of deoxyribonucleotides to the 3-end of single- and double-stranded DNA. This method has also been termed TUNEL (TdT-mediated dUTP-X nick end labeling). Alternatively, free 3-OH groups may be labeled using DNA polymerases by the template-dependent mechanism called nick translation. However, the TUNEL method is considered to be more sensitive and faster.
Sample material: Cells in suspension, cytospin and cell smear preparations, adherent cells grown on slides, and frozen and paraffin-embedded tissue sections.
Principle
The In Situ Cell Death Detection Kit, TMR red is based on the detection of single- and double-stranded DNA breaks that occur at the early stages of apoptosis.
Apoptotic cells are fixed and permeabilized. Subsequently, the cells are incubated with the TUNEL reaction mixture that contains TdT and TMR-dUTP. During this incubation period, TdT catalyzes the addition of TMR-dUTP at free 3-OH groups in single- and double-stranded DNA. After washing, the label incorporated at the damaged sites of the DNA is visualized by flow cytometry and/or fluorescence microscopy.
Kit for the detection and quantification of apoptotic cell death on a single-cell level by flow cytometry and fluorescence microscopy, and for double labeling with fluorescein-labeled cell markers (TMR red).
Specificity
The TUNEL reaction preferentially labels DNA strand breaks generated during apoptosis. This allows discrimination of apoptosis from necrosis and from primary DNA strand breaks induced by cytostatic drugs or irradiation.ApplicationIn Situ cell death detection kit, TMR red has been used in terminal deoxynucleotidyl transferase biotin–dUTP nick end labeling (TUNEL) assay. It has been used in apoptosis detection assay.
Precise, fast, and simple technique for detecting and quantitating apoptotic DNA fragmentation at the single-cell level in cells and tissues with a red fluorescent label for fluorescence microscopy and flow cytometry.
Packaging
1 kit containing 2 components.
Preparation Note
The TUNEL reaction mixture is prepared by mixing the Enzyme Solution and the Label Solution prior to use.
Working concentration: Enzyme concentration
The optimal enzyme concentration range from 0.5 to 5 U per assay. For a standard 50 µl PCR, we recommend using 2 U of the enzyme blend.
Working solution: Add total volume (50 µl) of Enzyme Solution to the remaining 450 µl Label Solution to obtain 500 µl TUNEL reaction mixture.
Mix well to equilibrate components.
Storage conditions (working solution): The TUNEL reaction mixture should be prepared immediately before use and should not be stored. Keep TUNEL reaction mixture on ice until use.Other NotesFor life science research only. Not for use in diagnostic procedures.Параметрыusage sufficient for 50 tests Quality Level 100, Торговая марка Roche shipped in dry ice storage temp. 20°C
Safety Informationpictograms GHS08,GHS09 signalword Danger hcodes H350i - H411 pcodes P201 - P273 - P280 - P308 + P313 - P391 - P501 RIDADR UN 1556 6.1 / PGIII WGK Germany WGK 3 Flash Point F does not flash Flash Point C does not flash -
11376497001
Insulin, human 11376497001
Empirical Formula (Hill Notation): C257H383N65O77S6 CAS Number: 11061-68-0 Molecular Weight: 5807.57 MDL number: MFCD00131380 General descriptionRecombinant Insulin, human, is produced in yeast and purified by standard chromatographic techniques. Insulin regulates blood glucose. In muscles and adipocytes it stimulates glucose intake and metabolism. It also controls the expression and various enzymes.
Specificity
Insulin, human, is active on most mammalian cells.ApplicationInsulin shows a broad range of activities on a variety of somatic cells. Recombinant human insulin can be used to stimulate growth and proliferation of cultured cells and to investigate insulin activity on sensitive cells used in research studies. It is also a component of serum-free media formulations for most primary cells and cell lines.
Biochem/physiol Actions
Two-chain polypeptide hormone produced by the β-cells of pancreatic islets. Its molecular weight is ~5800 Da. The α and β chains are joined by two interchain disulfide bonds. The α chain contains an intrachain disulfide bond. Insulin regulates the cellular uptake, utilization, and storage of glucose, amino acids, and fatty acids and inhibits the breakdown of glycogen, protein, and fat.Other NotesInChI: 1S/C257H383N65O77S6/c1-29-131(23)205(313-193(339)104-259)252(393)317-204(130(21)22)248(389)288-159(75-82-200(349)350)217(358)282-156(71-78-189(263)335)221(362)308-183-116-403-404-117-184-243(384)305-178(111-324)240(381)294-162(88-123(7)8)225(366)295-168(95-140-53-61-146(329)62-54-140)228(369)283-154(69-76-187(261)333)218(359)290-161(87-122(5)6)223(364)285-158(74-81-199(347)348)220(361)302-174(101-190(264)336)235(376)298-170(97-142-57-65-148(331)66-58-142)231(372)309-182(242(383)304-176(255(396)397)103-192(266)338)115-402-401-114-181(214(355)273-107-194(340)278-153(72-79-197(343)344)216(357)281-151(51-42-84-271-257(267)268)212(353)272-108-195(341)279-166(93-138-46-36-32-37-47-138)227(368)297-167(94-139-48-38-33-39-49-139)230(371)299-171(98-143-59-67-149(332)68-60-143)238(379)320-208(135(27)327)254(395)322-85-43-52-186(322)246(387)286-152(50-40-41-83-258)222(363)321-209(136(28)328)256(398)399)311-250(391)203(129(19)20)316-236(377)164(90-125(11)12)292-229(370)169(96-141-55-63-147(330)64-56-141)296-224(365)160(86-121(3)4)289-210(351)133(25)277-215(356)157(73-80-198(345)346)287-247(388)202(128(17)18)315-237(378)165(91-126(13)14)293-233(374)173(100-145-106-270-120-276-145)301-239(380)177(110-323)280-196(342)109-274-213(354)180(113-400-405-118-185(310-244(183)385)245(386)319-207(134(26)326)253(394)306-179(112-325)241(382)318-206(132(24)30-2)251(392)312-184)307-226(367)163(89-124(9)10)291-232(373)172(99-144-105-269-119-275-144)300-219(360)155(70-77-188(262)334)284-234(375)175(102-191(265)337)303-249(390)201(127(15)16)314-211(352)150(260)92-137-44-34-31-35-45-137/h31-39,44-49,53-68,105-106,119-136,150-186,201-209,323-332H,29-30,40-43,50-52,69-104,107-118,258-260H2,1-28H3,(H2,261,333)(H2,262,334)(H2,263,335)(H2,264,336)(H2,265,337)(H2,266,338)(H,269,275)(H,270,276)(H,272,353)(H,273,355)(H,274,354)(H,277,356)(H,278,340)(H,279,341)(H,280,342)(H,281,357)(H,282,358)(H,283,369)(H,284,375)(H,285,364)(H,286,387)(H,287,388)(H,288,389)(H,289,351)(H,290,359)(H,291,373)(H,292,370)(H,293,374)(H,294,381)(H,295,366)(H,296,365)(H,297,368)(H,298,376)(H,299,371)(H,300,360)(H,301,380)(H,302,361)(H,303,390)(H,304,383)(H,305,384)(H,306,394)(H,307,367)(H,308,362)(H,309,372)(H,310,385)(H,311,391)(H,312,392)(H,313,339)(H,314,352)(H,315,378)(H,316,377)(H,317,393)(H,318,382)(H,319,386)(H,320,379)(H,321,363)(H,343,344)(H,345,346)(H,347,348)(H,349,350)(H,396,397)(H,398,399)(H4,267,268,271)/t131-,132-,133-,134+,135+,136+,150-,151-,152-,153-,154-,155-,156-,157-,158-,159-,160-,161-,162-,163-,164-,165-,166-,167-,168-,169-,170-,171-,172-,173-,174-,175-,176-,177-,178-,179-,180-,181-,182-,183-,184-,185-,186-,201-,202-,203-,204-,205-,206-,207-,208-,209-/m0/s1
For life science research only. Not for use in diagnostic procedures.
Quality
Endotoxin level: <0.1EU/µg (LAL-test)
Note: 1 EU corresponds to 0.1ng
Sequence
A-chain: 21 AS, B-chain: 30 AS, connected by two disulfide bridges
Two polypeptide chains (A-chain: 21 amino acids, B-chain: 30 amino acids, connected by two disulfide bridges) identical to natural, human insulin.
Unit Definition
EC50 definition: The concentration of human insulin that is required to support half-maximal stimulation of cell proliferation (MTT cleavage) with 3T3 (A31) cells.
Physical form
Lyophilizate from a hydrochloric acid solution (pH 2.3) (crystalline insulin)
Preparation Note
Working concentration: 1-10 µg/ml
Recommended concentration for serum-free cell culture is 1-10 µg/ml.
The concentration of insulin required for stimulation of cell growth in almost all cases is extraordinarily high compared with the physiological concentration. Insulin may be mimicking insulin-like growth factors (IGFs, somatomedins) for some cell lines, and high insulin concentrations may be necessary to occupy receptors which have a high affinity for IGFs and a lower affinity for insulin.
Working solution: Dissolve Insulin, human, recombinant (100 mg or 500 mg), in sterile, double-dist. water (final concentration: 10 mg/ml).
Storage conditions (working solution): -15 to -25 °C
The reconstituted, undiluted solution is stable at -15 to -25 °C.
Note: Avoid repeated freezing and thawing.
Reconstitution
Reconstitute in sterile double-distilled water (final concentration 10 mg/ml), further dilution with PBS (phosphate buffered saline) or medium containing 1 mg/ml (0.1%) BSA (bovine serum albumin) [or HSA (human serum albumin)], or 1 to 10% serum.ПараметрыQuality Level 100 sterility non-sterile; 0.2 µm filtered assay >98% (SDS-PAGE) form lyophilized (clear, colorless solution after reconstitution) specific activity >26 units/mg protein (At least the same specific activity (EC<sub>50</sub>) compared to the indicated standard is guaranteed.) mol wt 5,700 Da packaging pkg of 100 mg Торговая марка Roche application(s) cell culture | mammalian: suitable impurities <0.1 EU/µg endotoxin (LAL test) solubility water: soluble UniProt accession no. P01308, shipped in wet ice storage temp. 2-8°C InChI key PBGKTOXHQIOBKM-FHFVDXKLSA-N Gene Information human ... INS(3630)
Safety InformationRIDADR NONH for all modes of transport WGK Germany nwg Flash Point F Not applicable Flash Point C Not applicable -
11074547001
Insulin-Transferrin-Sodium Selenite Supplement 11074547001
General descriptionThe insulin-transferrin-sodium selenite (ITS) supplement comprises three of the most crucial growth factors for many cell types. Insulin is a component of serum-free media formulations for all primary cells and cell lines. It stimulates cell growth, increases fatty acid and glycogen synthesis in a serum-free medium. Transferrin is an essential growth factor for many cell types. Selenium is very often necessary for optimal cell growth. ITS permits an appreciable reduction in serum requirements for cellular growth. ITS is also an antioxidant that supports the growth of in vivo derived (IVD) embryos. Insulin promotes embryonic growth and metabolism. The iron transport in the embryo is mediated by transferrin. Sodium selenite protects from oxidative damage and inhibits lipid peroxidation. ITS is useful in human chondrocytes monolayer culture.
The Insulin-Transferrin-Sodium Selenite Supplement contains the most essential growth-promoting components of serum-free culture media in an optimized ratio.
Source: Insulin- bovine pancreas; transferrin- human serum; Sodium selenite-syntheticApplicationThe Insulin-Transferrin-Sodium Selenite Supplement is used in the cell culture media as a basal growth-factor supplement to which other cell type-specific nutrients and growth factors are added.
Preparation Note
Working concentration: Insulin, 5 µg/ml, transferrin, 5 µg/ml, sodium selenite, 5 ng/ml (3.0 x 10-8 M)
Storage conditions (working solution): -15 to -25 °C.
Prepare appropriate aliquots and avoid repeated freezing and thawing.
Reconstitution
The insulin-transferrin-sodium selenite supplement should be reconstituted in 5 ml or 25 ml sterile double-dist. water (1000 x concentrated stock solution).
Further dilution with culture medium.Other NotesFor life science research only. Not for use in diagnostic procedures.ПараметрыQuality Level 100 biological source human serum (transferrin)","synthetic (sodium selenite) recombinant expressed in human cells sterility non-sterile; 0.2 µm filtered assay >98% (Transferrin, SDS-PAGE)",">99% (Insulin, HPLC)",">99% (Sodium selenite) form lyophilized packaging pkg of 50 mg (for 5 l medium) Торговая марка Roche application(s) cell culture | mammalian: suitable solubility water: miscible UniProt accession no. P01317,","P02786, shipped in wet ice storage temp. 2-8°C Gene Information bovine ... INS(280829)
human ... TFRC(7037)
Safety InformationRIDADR NONH for all modes of transport WGK Germany WGK 1 Flash Point F does not flash Flash Point C does not flash -
11040596001
Interferon-, human (hIFN-) 11040596001
General descriptionIFN-γ is produced by T-lymphocytes stimulated by antigen or by T-cell mitogens. Under nondenaturating conditions its molecular weight values range from 32,000 to 73,000 indicating that recombinant human IFN-γ exists as a dimer or higher oligomers.
Specificity
Human IFN-γ is effective on human cells, but not on mouse or rat cells.ApplicationInterferon-γ, human (hIFN-γ) has been used to stimulate mesenchymal stem cells (MSCs). It has also been used as a supplement Dulbecco′s modified Eagle′s medium/Ham′s medium for the cultivation of human colorectal carcinoma cell line T84 cells.
Interferon-γ, human (hIFN-γ), can be used to investigate IFN-γ activities in human cell systems.
Biochem/physiol Actions
Interferon-γ (IFN-γ) is an essential component of acquired immunity. A broad range of biological activities has been attributed to IFN-γ (e.g., the establishment of the antiviral state, immunoregulatory functions, antiproliferative effects and inhibition of cell growth). The anti-proliferative effects of IFN-γ are superior to those of either IFN-α or IFN-β. Growth inhibition is dependent on cell type, dose, and length of exposure. One of IFN-γ′s primary functions might be as an immunoregulatory agent: IFN-γ induces MHC antigens on many cells, Fc-receptors on monocytes and macrophages, IL-2 receptors on T-cells, enhances activity of macrophages, polymorphonuclear leukocytes, T-lymphocytes, and NK-cells (MAF), and is also involved in the regulation of B-cells.
A broad range of biological activities has been attributed to IFN-γ (e.g., the establishment of the antiviral state, immunoregulatory functions, antiproliferative effects and inhibition of cell growth). The anti-proliferative effects of IFN-γ are superior to those of either IFN-α or IFN-β. Growth inhibition is dependent on cell type, dose, and length of exposure. One of IFN-γs primary functions might be as an immunoregulatory agent: IFN-γ induces MHC antigens on many cells, Fc-receptors on monocytes and macrophages, IL-2 receptors on T-cells, enhances activity of macrophages, polymorphonuclear leukocytes, T-lymphocytes, and NK-cells (MAF), and is also involved in the regulation of B-cells.
Quality
The raw material used for this preparation was tested for HBs antigen and for the presence of antibodies to HIV-1, HIV-2, and HCV found to be negative.
Endotoxin level: <0.1EU/µg (LAL-test), <10EU/ml (LAL-test)
Note: 1EU corresponds to 0.1ng
Sequence
Chain Length 143 AA
The primary structure of recombinant human IFN-γ (143 amino acids) is identical to that of natural human IFN-γ (146 amino acids), however, recombinant IFN-γ has three amino acids less and is not glycosylated. Glycosylation is not essential for biological activity.
Unit Definition
EC50 definition: The amount of hIFN- that is required to produce equivalent antiviral activity to that expressed by 1 unit of the NIH IFN- reference standard (Gg 23– 901-530) (WISH cells-EMC virus/cytopathic effect) (1 unit equals 0.05 ng/ml).
Physical form
100,000U/ml or 1,000,000U/ml of Human IFN-γ in 0.1M phosphate buffer saline (PBS) (pH 7.0), 2.5% sucrose (w/v), and 2.5% human serum albumin (HSA) (w/v), filtered through a 0.2μm pore size membrane.
Preparation Note
Working solution: Solvent is recommended in double-distilled water.
Storage conditions (working solution): Stability in culture: Activity is present for at least 18 hours in culture medium.Other NotesFor life science research only. Not for use in diagnostic procedures.
EC 50 : <0.05ng/ml (hIFN- , NIH, reference standard, Gg 23–901-530)ПараметрыQuality Level 100 biological source human recombinant expressed in E. coli assay 95% (SDS-PAGE) form solution potency <0.05 ng/mL EC50 specific activity >2 x 10^7 U/mg mol wt 16,700 Da packaging pkg of 100,000 U (5 µg, 1 ml) Торговая марка Roche storage condition avoid repeated freeze/thaw cycles impurities <0.1 EU/µgtested (LAL test) UniProt accession no. P01579, shipped in dry ice storage temp. 20°C Gene Information human ... IFNG(3458)
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11276905001
Interferon-, mouse (mIFN-) 11276905001
General descriptionIFN-γ (interferon-γ) is produced by natural killer T cells, macrophages, Th1 CD4 T cells and activated CD8 T cells and the production increases with age.
Recombinant, Interferon-, mouse (mlFN-) is produced in E. coli and purified by standard chromatographic techniques.
Primary structure: One polypeptide chain (131 amino acids) identical to mouse, natural IFN- ? (133 amino acids), but lacking the last two amino acids at the carboxy-terminus and not glycosylated. Glycosylation is not essential for biological activity.
Specificity
Species specificity: mouseApplicationInterferon-γ, mouse (mIFN-γ) has been used in cell culture.
Recombinant mouse interferon-γ is a valuable tool for the study of IFN-γ (interferon-γ) actions in mouse model systems.
Biochem/physiol Actions
Interferon-γ (IFNγ) participates in cell-mediated immunity and has pro-inflammatory effects on immunocytes and target tissue. Its functions vary and is tissue specific. IFNγ induces T-cell-mediated immune response by increasing MHC-I expression on target cells and acts through IFNγ receptor, which is expressed on both immune and non-immune cells. IFNγ is associated with the development of colitis and Sjogren Syndrome.
Quality- Purity: >95% (SDS-PAGE)
- Endotoxin level: <0.1 EU/µg (LAL-test), <10 EU/ml (LAL-test)
Sequence
The primary structure of recombinant Interferon-γ, mouse (mIFN-γ) is identical to that of natural, mouse IFN-γ (one polypeptide chain, 131 amino acids), however, recombinant mIFN-γ lacks the last two amino acids at the C-terminus and is not glycosylated. Glycosylation is not essential for biological activity.
Chain Length 131 AA
Unit Definition
EC50 definition: One unit is defined as the amount of mlFN- that is required to produce equivalent antiviral activity to that expressed by 1 unit of the NIH IFN- reference standard (Gg 02-901-533). (L cells-EMC virus/cytopathic effect) (1 unit equals 0.2 ng/ml).
EC 50 definition : The amount of mIFN- that is required to produce equivalent antiviral activity to that expressed by 1 unit of the NIH IFN- reference standard (Gg 02–901-533) (L cells-EMC virus/cytopathic effect) (1 unit equals =0.2 ng/ml).
Physical form
Solution, filtered through 0.2 μm pore size membrane, 100 000 U/ml (20 μg/ml) in PBS (phosphate buffered saline) and BSA (bovine serum albumin),1 mg/ml [purity of BSA: 98%, endotoxin (LAL): < 1 EU/mg BSA].
Preparation Note
Working solution: Dilute the concentrated mlFN- solution (100,000 U/ml) with PBS or culture medium containing BSA, 1 mg/ml (0.1%) or serum, 1 to 10%.
Storage conditions (working solution): -15 to -25 °C
Store the solution in aliquots at -15 to -25 °C.
Avoid repeated freezing and thawing.Other NotesFor life science research only. Not for use in diagnostic procedures.
For life science research only. Not for use in diagnostic procedures.
1 EU corresponds to 0.1 ng.ПараметрыQuality Level 100, biological source mouse assay >95% (SDS-PAGE) form solution specific activity >5*10^6 U/mg ((mIFN- , NIH, reference standard, Gg 02–901-533, at least the same specific activity (EC 50 ) compared to the indicated standard is guaranteed.) packaging pkg of 100,000 U (20 µg, 1 ml) Торговая марка Roche shipped in dry ice storage temp. 20°C
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Interleukin-2, human (hIL-2)
Кат. номер 10799068001 11147528001 11011456001 General descriptionRecombinant Interleukin-2, human (hIL-2) is produced in E. coli and purified by standard chromatographic techniques. Solution in PBS and 1 mg/ml BSA, filtered through a 0.2 µm pore size membrane.
Specificity
Species specificity: Recombinant IL-2, human is effective on mouse and human cells.ApplicationRecombinant IL-2, human, allows:- The cultivation of human and murine IL-2 dependent T-cell lines and natural killer cell lines.
- The proliferation of mitogen-activated T-lymphocytes and natural killer cells.
- The establishment of human and murine thymocyte, splenocyte, or peripheral blood lymphocyte (PBL) derived T-cell lines.
- The generation of human and murine lymphokine-activated killer (LAK) cells (1-11).
- In vitro re-stimulation of lung cells obtained from mice.
Biochem/physiol Actions
Interlekin-2 (IL-2) is a potent pro-inflammatory cytokine. It plays a protective role in autoimmune chronic inflammation, when present in low amounts. It is involved in maintaining immune homeostasis where it controls the cross-talk between T regulatory and T effector cells. Studies in acute myelogenous leukemia (AML) mice models show that antibody based delivery of IL-2 to targets present in tumor-linked vasculature results in antileukemic activity.
Quality
Purity: >95% pure as determined by SDS-PAGE
Endotoxin level: <0.1 EU/µg (LAL-test), <10 EU/ml (LAL-test)
(1 EU corresponds to 0.1 ng)
Sequence
Chain Length 134 AA
The primary structure of recombinant, IL-2, human is identical to that of natural, human IL-2 (one polypeptide chain, 133 amino acids), however, recombinant IL-2 has an extra methionine at the amino-terminus (one polypeptide chain, 134 amino acids) and is not glycosylated. Glycosylation is not essential for biological activity.
Unit Definition
EC50 definition: The amount of hIL-2 that is required to support half-maximal stimulation of cell proliferation (XTT cleavage) with CTLL-2 cells (1 unit equals 0.5 ng).
Unit Conversion: 1 BM unit = 3.25 NBSB units (natural IL-2)1 BM unit = 3.25 NBSB units (natural IL-2)
Physical form
Solution in PBS and 1 mg/ml BSA, filtered through a 0.2 μm pore size membrane
Preparation Note
Working concentration: 10 -20 U/ml
Established IL-2-dependent T-cell lines usually require 10-20 U/ml. Add IL-2 to the freezing medium for IL-2 dependent cell lines.
Working solution: Dilute the concentrated IL-2 solution (200 U/ml or 10,000 U/ml) with PBS or culture medium containing 1 mg/ml BSA or HSA (0.1%) or 1 to 10% serum.
Storage conditions (working solution): -15 to -25 °C
It is recommended to store the solution in aliquots at -15 to -25 °C.
Note: Avoid repeated freezing and thawing.
Specific activity/EC 50 : >2 x 106 U/mg <0.5 ng/ml (hIL-2, NIBSC, 1st international standard, 86/504), at least the same specific activity (EC50 ) compared to the indicated standard is guaranteed (19, 20). Human, recombinant IL-2 has the same biological activity in vitro as compared to human, natural IL-2 (15, 1–4).
Recommended Method of Dilution: Dilute the concentrated IL-2 solution (200 U/ml or 10,000 U/ml) with PBS or culture medium containing 1 mg/ml BSA or HSA (0.1%) or 1–10% serum.
Interleukin-2 (IL-2, also known as T-Cell Growth Factor, TCGF) is a lymphokine which is produced by lectin- or antigen-activated T-cells and plays an important immunoregulatory role. This factor, or lymphokine, was first identified by its ability to promote the long-term in vitro proliferation of activated T cells. It also promotes the generation and proliferation of cytotoxic T-cells, natural killer (NK) cells, and lymphokine- activated killer (LAK) cells (1–11). Recombinant human IL-2 allows the cultivation of human and murine IL-2-dependent T-cell lines and natural killer cell lines, the proliferation of mitogenactivated T-lymphocytes and natural killer cells, the establishment of human and murine thymocyte-, splenocyte-, or peripheral blood lymphocyte (PBL)- derived T-cell lines, and the generation of human and murine lymphokine-activated killer (LAK) cells (1-11). Established IL-2-dependant T-cell lines usually require 10-20 U/ml. Add IL-2 to the freezing medium for IL-2 dependant cell lines.Other NotesFor life science research only. Not for use in diagnostic procedures.ПараметрыQuality Level 100 biological source human recombinant expressed in E. coli assay 95% (SDS-PAGE) form solution potency >2 x 10^6 units/mg EC50 mol wt (15,000 Da) packaging pkg of 10,000 U (10799068001 [5 µg, 50 ml])","pkg of 10,000 U (11011456001 [5 µg, 1 ml])","pkg of 50,000 U (11147528001 [25 µg, 5 ml]) Торговая марка Roche storage condition avoid repeated freeze/thaw cycles concentration 10-20U/ml impurities <0.1 EU/µg UniProt accession no. P60568, shipped in dry ice storage temp. 20°C Gene Information human ... IL2(3558)
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11271164001
Interleukin-2, mouse (mIL-2) 11271164001
General descriptionInterleukin-2 (IL-2, also known as T-cell growth factor, TCGF) is a lymphokine produced by lectin- or antigen-activated T-cells. It plays an important immunoregulatory role. This factor was first identified by its ability to promote the long-term in vitro proliferation of activated T-cells. It also promotes the generation and proliferation of cytotoxic T-cells, natural killer (NK) cells, and lymphokine-activated killer (LAK) cells. IL-2 also induces other lymphokines such as interferon-γ and B-cell growth factor (BCGF-1).
Recombinant Interleukin-2, mouse (mIL-2), is produced in E. coli and purified by standard chromatographic methods.
Contents: 10,000 U/ml solution in PBS and 1 mg/ml BSA, filtered through a 0.2 µm pore size membrane
Specificity
Mouse IL-2 is effective on mouse cells, but not on human cells.ApplicationRecombinant murine IL-2 is produced in E. coli and exhibits the following:- Supports the growth of murine CTLL cells (murine T cell line), but not that of human T-cells.
- Strongly inhibits the binding of recombinant human IL-2 to murine responder cells.
- Weakly inhibits the binding to human responder cells.
- Shares identical biological and immunological activities with human IL-2.
- Is a convenient tool for extensive studies of the pharmacological and physiological activities of IL-2 in murine models.
- used for the stimulation of murine T-cells
Quality
Endotoxin level: <0.1 EU/µg (LAL-test), <10 EU/ml (LAL-test)
Sequence
One polypeptide chain (149 amino acids), identical to natural mouse IL-2, but not glycosylated. Glycosylation is not essential for biological activity.
Chain Length 149 AA
Unit Definition
EC50 definition: The amount of mIL-2 that is required to support half-maximal stimulation of cell proliferation (XTT cleavage) with CTLL-2 cells (1 unit equals 0.5 ng).
Preparation Note
Working solution: Dilute the concentrated IL-2 solution (10,000 U/ml) with PBS or culture medium containing 1 mg/ml BSA [or HSA (human serum alburnin)], or 1 to 10% serum (v/v).
Storage conditions (working solution): -15 to -25 °C
Note: Avoid repeated freezing and thawing.Other NotesFor life science research only. Not for use in diagnostic procedures.
Specific activity/EC 50: >2 x 106 U/mg, <0.5 ng/ml (hIL-2, NIBSC, 1st international standard, 86/504), at least the same specific activity (EC50) compared to the indicated standard is guaranteed.
Note: The biological activity of this product may vary in different in vitro applications. Determine the optimal concentration range for specific applications.
EC 50 definition/Unit definition: The amount of mIL-2 that is required to support half-maximal stimulation of cell proliferation (XTT cleavage) with CTLL-2 cells (1 unit equals =0.5 ng).ПараметрыQuality Level 100 biological source mouse recombinant expressed in E. coli assay >95% (SDS-PAGE) form solution potency <0.5 ng/mL EC50 mol wt 17,200 Da packaging pkg of 10,000 U (5 µg, 1 ml) Торговая марка Roche storage condition avoid repeated freeze/thaw cycles impurities <0.1 EU/µgtested (LAL test) UniProt accession no. P04351, shipped in dry ice storage temp. 20°C Gene Information mouse ... Il2(16183)
Safety InformationRIDADR NONH for all modes of transport WGK Germany nwg Flash Point F No data available Flash Point C No data available -
11138600001
Interleukin-6, human (hIL-6) 11138600001
General descriptionInterleukin-6, human (hIL-6), is produced in E. coli and purified by standard chromatographic techniques.
Specificity
Recombinant IL-6, human is effective on mouse and human cells.
IL-6, a 26 kD protein, also known as B-cell stimulatory factor 2 (BSF-2, BCDF), interferon-2 (IFN-2), hybridoma/plasmacytoma growth factor (H(P)GF), and hepatocyte-stimulating factor is produced by many cell types (e.g., T-cells, monocytes, fibroblasts, endothelial cells). IL-6 induces the terminal maturation of activated B-cells into antibody-producing cells (B-cell differentiation activity. The hepatic acute phase response is induced by IL-6 (hepatocyte-stimulating activity). It was shown that IL-6 and IL-3 act synergistically to support the proliferation of hematopoietic stem cells (hemapoietic activity), however, IL-6 does not have antiviral activity, although it has been called IFN-2. High affinity receptors for IL-6 are found on many cell types.ApplicationRecombinant IL-6, human, is a growth factor for murine and human B-cells. It can be used to replace feeder cells in the preparation of murine and human hybridomas.
Biochem/physiol Actions
Interleukin-6 participates in several pleiotropic functions in various types of cells through its specific receptor (IL-6-R). It has been suggested that interleukin-6 may participate in the interaction between neuroendocrine and immune system by activating adrenocorticotropic hormone secretion. It has also been reported that IL-6 stimulates the bonding of receptor (IL-6-R) with a non-ligand-binding membrane glycoprotein, gp130.
Quality
Purity: 95% pure as determined by HPLC or SDS-PAGE [Endotoxin level: <0.1 EU/µg (LAL-test), <10 EU/ml (LAL-test).
Note: 1 EU corresponds to 0.1 ng
Sequence
Chain Length 184 AA
The primary structure of recombinant human IL-6 is identical to that of natural human IL-6 (one polypeptide chain, 184 amino acids), however, it contains an altered sequence within the first 15 amino-terminal amino acids.
Unit Definition
EC50 definition: The amount of hIL-6 that is required to support half-maximal stimulation of cell proliferation (MTT cleavage) with 7TD1 cells (1 unit equals <0.01 ng).
Physical form
Solution, filtered through 0.2 μm pore size membrane.
Preparation Note
Working concentration: 10 to 100 U/ml (0.1 1 ng/ml)
10 to 100 U/ml (0.1 to 1 ng/ml)
A concentration of 100 U/ml (1 ng/ml) is recommended for the preparation of B-cell hybridomas
Working solution: Dilute the concentrated IL-6 solution (200,000 U/ml) with PBS or culture medium containing 1 mg/ml BSA [or HSA (human serum albumin)] or 1 to 10% serum.
Storage conditions (working solution): -15 to -25 °C
Store the solution in aliquots at -15 to -25 °C.
Note: Avoid repeated freezing and thawing.
Reconstitution
Dilute the concentrated IL-6 solution (200,000 U/ml) with PBS or culture medium containing 1 mg/ml BSA [or HSA (human serum albumin)] or 1 to 10% serum.
Analysis Note
Specific activity/EC50: >1 x 108 U/mg, <0.01 ng/ml (hIL-6, NIBSC, interim standard, 88/514), at least the same specific activity (EC50) compared to the indicated standard is guaranteed.Other NotesFor life science research only. Not for use in diagnostic procedures.ПараметрыQuality Level 100 biological source human recombinant expressed in E. coli assay >98% (SDS-PAGE) form solution potency <0.01 ng/mL EC50 mol wt 20,600 Da packaging pkg of 200,000 U (2 µg, 1 ml) Торговая марка Roche storage condition avoid repeated freeze/thaw cycles impurities <0.1 EU/µgtested (LAL test) UniProt accession no. P05231, shipped in dry ice storage temp. 20°C Gene Information human ... IL6(3569)
Safety InformationRIDADR NONH for all modes of transport WGK Germany nwg Flash Point F does not flash Flash Point C does not flash -
Isopropyl--D-thiogalactoside
Empirical Formula (Hill Notation): C9H18O5S CAS Number: 367-93-1 Molecular Weight: 238.30 Beilstein/REAXYS Number: 4631 MDL number: MFCD00063273 PubChem Substance ID: 329815691
Кат. номер 11411446001 10724815001 General descriptionIsopropyl--D-thiogalactoside (IPTG) is a chemical analog of galactose that cannot be cleaved by -galactosidase. IPTG functions by binding to the lacI repressor and altering its conformation, which prevents the repression of the -galactosidase coding gene lacZ.ApplicationIsopropyl--D-thiogalactoside (IPTG) is commonly used in cloning procedures that require induction of -galactosidase activity. It is used in conjunction with X-Gal or Bluo-Gal in blue-white selection of recombinant bacterial colonies that induce expression of the lac operon in Escherichia coli. It has been used for the expression of recombinant genes in E. coli.
IPTG is commonly used in cloning procedures that require induction of -galactosidase activity. It is used in conjunction with X-Gal or Bluo-Gal in blue-white selection of recombinant bacterial colonies that induce expression of the lac operon in Escherichia coli. IPTG functions by binding to the lacI repressor and altering its conformation, which prevents the repression of the -galactosidase coding gene lacZ.Other NotesNon-metabolizable galactose analog.
For life science research only. Not for use in diagnostic procedures.
The amount of labeled DNA depends on:- amount of template DNA
- purity of template DNA
- conformation of template DNA
- average size of labeled fragment
Specifications
Assay Time: 50minutes
Specific Activity: The described standard assay will results in a specific activity of 1.8 x 109 dpm/µg, corresponding to 65% incorporation with different substrate DNAs in 30minutes. When varying the ratio of template DNA to labeled dNTP, similar incorporation rates, but different levels of specific activity of the labeled probe are obtained.
Preparation Note
Sample Material
10 ng to 3µg template DNA, linearized and at least 100 to 200bp long
Working concentration: Final concentration should be 0.2 mM.
Storage conditions (working solution): Stock solutions of the product in water (e.g., 1 M IPTG) stored at -15 to -25 °C, are stable up to six months.Параметрыmol wt 238.3 packaging pkg of 1 g (10724815001), pkg of 5 g (11411446001) Торговая марка Roche shipped in wet ice storage temp. 2-8°C SMILES string CC(C)S[C@@H]1OC(CO)[C@H](O)C(O)[C@H]1O InChI 1S/C9H18O5S/c1-4(2)15-9-8(13)7(12)6(11)5(3-10)14-9/h4-13H,3H2,1-2H3/t5-,6+,7+,8-,9+/m1/s1 InChI key BPHPUYQFMNQIOC-NXRLNHOXSA-N
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11493027001
IsoStrip™ Mouse Monoclonal Antibody Isotyping Kit 11493027001
General descriptionIsoStrip Mouse Monoclonal Antibody Isotyping Kit is a simple, three-step kit for the rapid characterization of mouse monoclonal antibodies.
Contents- Development Tubes, contain lyophilized latex beads, precoated with anti-mouse-Ig antibodies
- Isotyping Strips, precoated with subclass- and light-chain-specific anti-mouse-Ig antibodies
Specificity
Anti-mouse antibodies, bound to the latex beads, will react with any mouse monoclonal antibody regardless of its isotype.Goat anti-mouse antibodies, immobilized to the strip, specifically bind to each of the common mouse antibody isotypes (IgG1, IgG2a, IgG2b, IgG3, IgM, and IgA), and to the light chains.
Specificity for all specific antibodies beside IgML at 1.0 µg/ml: 2
IgM Lambda, at 1.0 µg/ml: 1ApplicationThe kit is used for the rapid identification of class, subclass, and light-chain types of mouse monoclonal antibodies (immunoglobulins).
Features and Benefits- Reliable: Yields comparable results to the standard ELISA assay
- Fast: 5 minutes from start to finish
- Easy: Only 3 steps
- Economical: No equipment or additional reagents required
- Highly specific: No crossreactivity with FBS
Packaging
1 kit containing 2 components.
Specifications
Assay time: 5 minutes
Sample material: Hybridoma supernatant, purified mouse monoclonal antibodies, ascites
Preparation Note
Working solution: Dilute Sample- Remove the desired number of isotyping strips from the canister. Remove the caps from an equal number of development tubes.
The tubes may be labeled with a pencil or felt-tipped lab marker for easy identification. - Dilute a sample containing the mouse monoclonal antibody in 1% BSA/ phosphate-buffered saline (PBS), pH 7.2 to 7.6.
Culture supernatant samples should be diluted 1:10 to 1:100. Ascites samples should be diluted 1:20,000. These are recommended dilutions and may vary depending on the concentration of antibody in your sample. In our experience, a monoclonal antibody
concentration of 0.1 to 1 g/ml of diluted sample gives the best results. 150 ml of this diluted sample will be added to the development tube.
Other NotesFor life science research only. Not for use in diagnostic procedures.Legal InformationIsoStrip is a trademark of RocheПараметрыusage sufficient for 10 tests Quality Level 100, Торговая марка Roche shipped in wet ice storage temp. 2-8°C
Safety InformationRIDADR NONH for all modes of transport WGK Germany WGK 1 Flash Point F does not flash Flash Point C does not flash -
KAPA2G Fast Multiplex Mix
NACRES: NA.55
Кат. номер KK5802 KK5801 General descriptionKAPA2G Fast Multiplex PCR Kits contain a second-generation (2G) enzyme derived through a process of directed evolution. KAPA2G FAST HotStart® DNA Polymerase is an antibody-mediated hot start formulation engineered for higher processivity and speed, offering significantly faster extension rates than wild-type Taq DNA polymerase. In addition to speed, KAPA2G Fast provides higher yields and sensitivity than competitor enzymes, allowing for more uniform multiplexed PCR.
KAPA2G Fast HotStart PCR Kits are designed for fast PCR, in which total reaction times are 20–70% shorter than those of conventional PCR assays performed with wild-type Taq DNA polymerase. This can be achieved without sacrificing reaction performance and does not require specialized PCR consumables or thermocyclers.ApplicationKAPA2G Fast Multiplex Mix has been used for:- Typing of transgenic organisms
- Amplification of microsatellites
- Typing and detection of pathogens
- Amplification of multiple DNA fragments for single nucleotide polymorphism (SNP) genotyping
- Semi-quantitative PCR
- 3-plex PCR
Biochem/physiol Actions
DNA fragments generated with KAPA2G Fast DNA Polymerase have the same characteristics as DNA fragments generated with wild-type Taq DNA polymerase, and may be used for sequencing, restriction enzyme digestion and cloning. Like wild-type Taq, KAPA2G Fast has 53 polymerase and 53 exonuclease activities, but no 35 exonuclease (proofreading) activity. The fidelity of KAPA2G Fast is similar to that of wild-type Taq; it has an error rate of approximately 1 error per 1.7 x 105 nucleotides incorporated. PCR products generated with KAPA2G Fast are 3-dA-tailed and may be cloned into TA cloning vectors.
Features and Benefits
Improve sensitivity, specificity, and yields- Uniform representation of all amplicons
- Successful multiplex PCR with difficult, GC-rich targets
Increase speed without compromising performance- 60% reduction in cycling time
- Extension times as low as 15 seconds
Quick Notes:- KAPA2G Fast Multiplex Mix contains the engineered KAPA2G Fast HotStart DNA Polymerase, for fastand efficient multiplex PCR.
- The KAPA2G Fast Multiplex Mix contains a buffer optimized for multiplex PCR, with 0.2 mM of each dNTP and 3 mM MgCl2 (at 1X).
- Use 0.2 µM of each primer, and 10–100 ng of template DNA per reaction.
- Anneal at 60°C for 30 seconds.
- Perform extension for 15 sec, and increase for longer amplicons, and/or highly multiplexed reactions.
Quality
Each batch of KAPA2G Fast HotStart DNA Polymerase is confirmed to contain <2% contaminating protein (AgilentProtein 230 Assay). KAPA2G Fast Multiplex PCR Kits are subjected to stringent quality control tests, are free of contaminating exo- and endonuclease activity, and meet strict requirements with respect to DNA contaminationlevels.
Preparation Note
Always ensure that the product has been fully thawed and mixed before use. Reagents may be stored at 4°C for short-term use (up to 1 month). Return to -20°C for long-term storage. Provided that the ReadyMix has been handled carefully and not contaminated, the kit is not expected to be compromised if left (unintentionally) at room temperature for a short period of time (up to 3 days). Long-term storage at room temperature and 4°C is not recommended. Please note that reagents stored at temperatures above -20°C are more prone to degradation when contaminated during use, and therefore storage at such temperatures is at the users own risk.Other NotesFor Research Use Only. Not for use in diagnostic procedures.Legal InformationHOTSTART is a registered trademark of Molecular BioProducts, Inc.Параметрыshelf life ≤12 mo. Quality Level 100, packaging kit of 1.25 mL (100 x 25 μL rxn; KK5801), kit of 6.25 mL (500 x 25 μL rxn; KK5802) Торговая марка Roche concentration 2 x application(s) PCR: suitable shipped in dry ice storage temp. −20°C
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10104531001
Klenow Enzyme 10104531001
General descriptionKlenow enzyme is the large, C-terminal fragment (Mr 76,000) of E.coli DNA polymerase I, which can be obtained by subtilisin treatment of intact DNA polymerase I. It retains the 53 polymerase and the 35 exonuclease activities of intact DNA polymerase I, but lacks the 53 exonuclease activity of the native enzyme. The enzyme catalyzes the addition of mononucleotides from deoxynucleoside-5-triphosphates to the 3-hydroxyl terminus of a primer/template DNA. This property is used to synthesize DNA complementary to single-stranded DNA templates.
Klenow Enzyme is formed by subtilisin treatment of DNA polymerase I.ApplicationKlenow Enzyme has been used in blunt end ligation of recyclin-1 (RCY1) gene into pETDUET-GSTHis6-S Tag vector.
Quality
Function test: Sequencing Grade Klenow Enzyme is tested in sequencing reactions according to Sanger. The test is conducted with 1 unit of enzyme and the M13 sequencing system. Sequencing gels are examined by autoradiography. The readable sequence must be 250 bases.
Absence of contaminants: Sequencing Grade Klenow Enzyme is tested on various DNA substrates to ensure the absence of nonspecific endonucleases and exonucleases.
Unit Definition
One unit is the enzyme activity which incorporates 10 nmol of total nucleotides into an acid-precipitable fraction in 30 minutes under the following assay conditions:
Incubation Buffer
130 mM Potassium phosphate, 6.5 mM MgCI2, 33 µM 14C-dTTP, poly[d(A-T)], 0.833 A260/ml; dATP, 33 µM; 1 mM dithioerythritol; bovine serum albumin, 0.032 mg/ml; pH 7.4.
Incubation Procedure
0.05 to 0.25 units of enzyme are incubated for 30 minutes at +37 °C in a total volume of 300 µl incubation buffer.
Volume Activity: 1,000 units/ml, or 5,000 units/ml respectively, according to Richardson (+37 °C, poly [d(A-T)] as primer).
Physical form
Solution of enzyme, 5U/µl, in storage buffer:
50 mM Potassium phosphate, 1 mM dithioerythritol, glycerol, 50% (v/v); pH 7.0 (4 °C)Other NotesFor life science research only. Not for use in diagnostic procedures.ПараметрыQuality Level 100, form solution specific activity >5000 units/mg protein mol wt Mr 75 kDa packaging pkg of 250 U (5 x 103 U/ml) Торговая марка Roche shipped in dry ice storage temp. 20°C (15°C to 25°C)
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11242008001
L-Carnitine 11242008001
General descriptionEnzymatic UV-Test with a linearity in the range of 5.6 - 112 μM L-carnitine in the test solution.ApplicationDetermination of L-carnitine in seminal plasma, serum, or urine in life science research applications.
Biochem/physiol Actions
L-carnitine is an abundant amino acid present in red meat. Its trimethylamine structure has similarity to that of choline. The major source of this amino acid is by dietary ingestion. In mammals, it is also endogenously produced from lysine. It participates in the transport of fatty acids into the mitochondrial compartment. It produces trimethylamine-N-oxide (TMAO) and promotes atherosclerosis.
Packaging
1 kit containing 6 components
Principle
L-Carnitine is acetylated to acetyl carnitine by acetyl coenzyme A (acetyl CoA) in the presence of the enzyme carnitine acetyl transferase (CAT). The resulting coenzyme A (CoA) is acetylated back to acetyl CoA in the presence of adenosine-5-triphosphate (ATP) and acetate, catalyzed by the enzyme acetyl CoA synthetase (ACS). This results in the formation of adenosine-5-monophosphate (AMP) and inorganic pyrophosphate (PPi). In the presence of ATP, supported by myokinase (MK), AMP forms twice the amount of adenosine-5-diphosphate (ADP). This is converted in the following reaction with phosphoenol pyruvate (PEP) in the presence of pyruvate kinase (PK). The pyruvate formed is reduced to L-lactate by reduced nicotinamide adenine dinucleotide (NADH) in the presence of lactate dehydrogenase (LDH). The amount of NADH consumed during the reaction is equivalent to half the amount of L-carnitine. NADH is measured spectroscopically at 340 nm.
Preparation Note
Storage conditions (working solution): Note:- After reconstitution, the solution 1 is stable for 4 days at 2 to 8 °C.
- Bring solution 1 to 15 to 25 °C prior to use.
- After reconstitution, the suspension 2 is stable for 3 months at 2 to 8 °C.
Other NotesFor life science research only. Not for use in diagnostic procedures.Параметрыusage sufficient for ~3 x 10 determinations (test-combination), sufficient for ~3 x 10 tests Торговая марка Roche shipped in wet ice storage temp. 2-8°C
Safety Informationpictograms GHS05 signalword Danger hcodes H318 - H412 pcodes P273 - P280 - P305 + P351 + P338 + P310 - P501 RIDADR NONH for all modes of transport WGK Germany WGK 2 Flash Point F does not flash Flash Point C does not flash -
L-Lactate Dehydrogenase (L-LDH)
Enzyme Commission (EC) Number: 1.1.1.27 ( BRENDA | IUBMB )
Кат. номер 10127230001 10127884001 10127876001 General descriptionL-lactate:NAD+ oxidoreductase.
L-lactate dehydrogenase catalyzes the reversible reduction of pyruvate to L-lactate.
Specificity
L()-Lactate dehydrogenase (LDH) is specific for L()-lactate (relative rate = 100). It does not react with D(-)-lactate.LDH will also slowly oxidize glyoxylate (oxoacetate), glycerate and 3-halogen derivatives of L-lactate (rate with each, 1); 3-phosphoglycerate does not react.LDH will reduce pyruvate (2-oxopropanate; relative rate = 100) and other 2-oxoacids (e.g., 2-oxobutyrate), with rates decreasing rapidly with increasing chain length of the acid. Heart LDH reacts with 2-oxobutyrate at a greater rate than does skeletal muscle LDH.LDH will also oxidize 2,4-diketoacids (relative rate = 10). The enzyme is realtively specific for NAD(H) (relative rate = 100); NADP(H) is utilized much less efficiently (relative rate < 1). Representative Kms for (hog) skeletal muscle LDH (pH 7.5) are lactate, 8.3 mM; pyruvate, 0.12 mM; NAD, 0.10 mM; NADH, 0.012 mM.Representative Kms for (pig) heart LDH (pH 7.5) are lactate, 3.3 mM; pyruvate, 0.15 mM; NAD, 0.07 mM;NADH, 0.011 mM. In general, the concentration of pyruvate (reduction reaction) or of L-lactate (oxidation reaction) necessary to obtain a maximal rate with LDH is lower for preparations from heart tissue than for the preparations from skeletal muscle.ApplicationReduction of α-ketoacids to α-hydroxycarboxylic acids, or reverse reaction.
Sequence
LDH is a tetramer (MW = approx. 140,000 D, rabbit muscle or pig heart LDH). In mammals, there are two types of LDH subunits, M and H, with similar molecular weight (subunit MW 36,500 D) but differing amino acid composition.
Therefore, 5 electrophoretically distinguishable LDH isoenzymes, LDH-1 through LDH-5, of differing subunit composition, are found in mammals.
The molecular weight of all LDH isoenzymes is approximately the same.
– LDH-1 (H4), LDH-2 (H3M): predominant components of heart LDH
– LDH-3 (H2M2): principal component of LDH from lymphatic tissue
– LDH-4 (HM3), LDH-5 (M4): predominate in skeletal muscle or liver LDH
Note: The M subunit of LDH used to be designated the A subunit; the H subunit was the B subunit. Under the old designation, LDH-1 isoenzyme was LDH-B4.
Unit Definition
One unit (U) lactate dehydrogenase will reduce 1 mol of pyruvate to L-lactate in 1 minute at +25 °C and pH 7.0.
Note: For preparations H and I, the activity is defined at +30 °C, pH 7.8. The above assay consumes 1 mol of NADH per mol of pyruvate reduced.
Unit Conversion: For the reduction reaction, pyruvate as substrate:
1 U (+25 °C) 1.5 U (+37 °C) [hog muscle LDH in glycerol].
1 U (+25 °C) 1.8 U (+37 °C) [hog muscle LDH in amm onium sulfate].
1 U (+25 °C) 1.1 U (+30 °C) 2.0 U (+37 °C) [rabbit muscle LDH].
1 U (+25 °C) 1.4 U(+30 °C) 2.5 U (+37 °C) [pig heart LDH].
1 U (+25 °C) 2.5 U (+37 °C) [beef heart LDH].
Physical form
Suspension in 3.2 M ammonium sulfate solution, pH approximately 7
Preparation Note
Activator: 2-amino-2-methyl-1-propanol, diethanolamine, fluoride and heparin (oxidation reaction). Sodium sulfite (Na2SO3) will protect the enzyme from the effects of thiol-attacking reagents. Mercaptans can reverse the inhibitory effects of thiol-attacking reagents.
Analysis Note
Absorbance:The purified enzyme from beef heart has an absorbance of 14.9 (10 mg/ml, 280 nm).Other NotesFor life science research only. Not for use in diagnostic procedures.Параметрыform suspension Quality Level 100, specific activity ~550 units/mg protein (at 25 °C (1,100 U/mg at 37 °C) with pyruvate as the substrate.) packaging pkg of 10 mL (10127884001 [100 mg]), pkg of 2 mL (10127230001 [10 mg]), pkg of 5 mL (10127876001 [25 mg]) Торговая марка Roche shipped in wet ice storage temp. 2-8°C
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10107085001
L-Lactate Dehydrogenase (L-LDH) 10107085001
Enzyme Commission (EC) Number: 1.1.1.27 ( BRENDA | IUBMB ) General descriptionL-Lactate Dehydrogenase (L-LDH) is an enzyme which is involved in the glycolytic pathway. It is believed to consist of five isotypes, LDH1 to LDH5. It also has a major role in carbohydrate metabolism of human malaria parasites.ApplicationReduction of α-ketoacids to α-hydroxycarboxylic acids or reverse reaction.
It was used in determination of D-Lactate.
Physical form
Suspension in 3.2 M ammonium sulfate solution, pH approximately 6.5Other NotesFor life science research only. Not for use in diagnostic procedures.ПараметрыQuality Level 100, form suspension specific activity ~550 units/mg protein (at 25 °C (1,000 U/mg at 37 °C) with pyruvate as the substrate.) packaging pkg of 10 mL (100 mg) Торговая марка Roche
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L-Malate Dehydrogenase (L-MDH)
Enzyme Commission (EC) Number: 1.1.1.37 ( BRENDA | IUBMB )
Кат. номер 10127256001 10127914001 10127248001 General descriptionL-malate:NAD+ oxidoreductase
The L-malate dehydrogenase enzyme is a nuclear gene product that is synthesized with a 24-residue amino-terminal signal peptide. This peptide is proteolytically cleaved during the translocation of the enzyme to the mitochondrial matrix.ApplicationThe enzyme L-malate dehydrogenase from pig heart has been used to measure PEPCK (phosphoenolpyruvate carboxykinase) activity. The oxaloacetate, produced by PEPCK, is reduced to malate via the oxidation of NADH, which in turn is measured at 340 nm using a spectrophotometer. The enzyme has also been used to measure oxaloacetate by measuring the reduction in NADH spectroscopically at 340 nm.
Biochem/physiol Actions
The enzyme L-malate dehydrogenase from pig heart catalyzes the oxidation of L-malate to oxaloacetate. The enzyme is an NAD-dependent mitochondrial dehydrogenase that functions in the tricarboxylic acid cycle. It is a component of the malate-aspartate shuttle that transports reducing equivalents across the inner mitochondrial membrane in the form of malate.
Quality
Contaminants: <0.002% GOT, <0.01% fumarase and LDH, each luM each, <0.02% HK, <0.002% PGI
Physical form
Suspension in 3.2 M ammonium sulfate solution, pH approximately 6
Solution in 50% glycerol (v/v), pH approximately 7
Preparation Note
Activator: – phosphate
– arsenate
– Mg2+
– Zn2+Other NotesFor life science research only. Not for use in diagnostic procedures.ПараметрыQuality Level 100, form solution, suspension specific activity ~1200 units/mg protein (At 25 °C with oxaloacetate as the substrate.) packaging pkg of 1 mL (10127248001 solution), pkg of 1 mL (10127256001 suspension), pkg of 5 mL (10127914001suspension) Торговая марка Roche optimum pH 7.4-7.5(reduction of oxaloacetate), 9.2-9.5(oxidation of malate) shipped in wet ice storage temp. 2-8°C
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11243217001
Laminin 11243217001
CAS Number: 114956-81-9 MDL number: MFCD00081739 General descriptionLaminin is purified as laminin-nidogen complex from mouse Engelbreth-Holm-Swarm (EHS) sarcoma. Laminin is a heterotrimer made from , and chain subunits. It is a glycoprotein which is required for the structural scaffolding of basement membranes. Laminin supports growth and differentiation of many cell types including epithelial, endothelial, neural, muscle and liver cells.
Biological activity: Tested for the promotion of adherence of HT-1080 cells.
Species specificity: Active on most mammalian cells.ApplicationLaminin has been demonstrated to promote the attachment and growth of a variety of cells; used for the coating of culture dishes. It has been used for cell adhesion assay. It is recommended for use as a cell culture substratum at 1-2μg/cm2. The optimal concentration does depend on cell type as well as the application or research objectives.
Laminin supports growth and differentiation of many cell types including epithelial, endothelial, neural, muscle and liver cells. It is recommended for use as a cell culture substratum at 1-2 µg/cm2. The optimal concentration does depend on cell type as well as the application or research objectives.
Quality
Purity: 90% (SDS-PAGE) (as laminin nidogen-complex, 1:1)
Sequence
Mouse laminin is composed of three polypeptide chains (A: 400 kD, B1: 230 kD and B2: 220 kD), connected by disulphide bridges, and is glycosylated.
Physical form
Solution, 0.5 mg/ml laminin in 0.15 M NaCl, 2 mM EDTA, 0.05 M Tris-HCl, pH 7.4, sterile
Solution, filtered through 0.2 ?m pore size membrane
Preparation Note
Working concentration: 2 to 5 µg/cm2
2 to 5 µg/cm2 for the coating of cell culture vessels
Storage conditions (working solution): 2 to 8 °C
The undiluted solution is stable at 2 to 8 °C for at least 3 months.
The solution should be thawed carefully at 2 to 8 °C or on ice.Other NotesFor life science research only. Not for use in diagnostic procedures.ПараметрыQuality Level 100 biological source mouse (Engelbreth-Holm-Swarm (EHS) sarcoma) sterility sterile; sterile-filtered assay 90% (SDS-PAGE) form solution mol wt 900 kDa packaging pkg of 1 mg (2 ml) Торговая марка Roche application(s) cell culture | mammalian: suitable UniProt accession no. Q61292, shipped in dry ice storage temp. 20°C Gene Information mouse ... Lamb2(16779)
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Leupeptin
Empirical Formula (Hill Notation): C20H38N6O4 · 0.5H2O4S Linear Formula: C20H38N6O4 · 1/2H2SO4 CAS Number: 103476-89-7 Molecular Weight: 475.59 MDL number: MFCD00037012 PubChem Substance ID: 329817679
Кат. номер 11017101001 11529048001 11034626001 11017128001
Specificity
Leupeptin inhibits serine and thiol proteases such as trypsin, plasmin, proteinase K, kallikrein, papain, thrombin, and cathepsin A and B.
Not affected are -, -, - and - chymotrypsin, pepsin, cathepsin D, elastase, renin, and thermolysin.ApplicationLeupeptin as well as other protease inhibitors like antipain, chymostatin, pepstatin, and phosphoramidon are useful for the protection of proteins during their isolation from tissues or membranes. Leupeptin can be removed from the reaction by dialysis.
Note: To check other protease inhibitors, try our Protease Inhibitor Set including Antipain Dihydrochloride, Aprotinin, Bestatin, Chymostatin, E-64, EDTA-Na2, Leupeptin, Pefabloc SC, Pepstatin, and Phosphoramidon.
Biochem/physiol Actions
Inhibitor of serine and cysteine proteases. Inhibits plasmin, trypsin, papain, calpain, and cathepsin B. Does not inhibit pepsin, cathepsins A and D, thrombin, or α-chymotrypsin. Effective concentration 10-100 μM. There have been numerous studies using leupeptin to protect against hearing loss caused by acoustic overstimulation or the ototoxic antibiotic gentamicin. (Loss of cochlear hair cells is believed to be mediated by calpain.)
Features and Benefits
Contents
Synthetic, white powder
Quality
Function test: Performance controlled with trypsin.
Preparation Note
Working concentration: 0.5 to 5 µg/ml (1 to 20 µM)
Comparison of working concentrations of pefabloc with leupeptin and PMSF is in files.
Working solution: Solvent is recommended in distilled water.
Highly soluble in water (1 mg/ml), methanol, ethanol, acetic acid, dimethyl formamide and dimethyl sulfoxide.
Poorly soluble in acetone, chloroform, ethyl ether and n-hexane.
Storage conditions (working solution): -15 to -25 °C
In aqueous solution leupeptin is stable for 1 month at 2 to 8 °C or for at least 6 months at - 15 to -25 °C, stored under nitrogen. For best results, freeze the dissolved inhibitor in aliquots and avoid repeated thawing.
Reconstitution
Highly soluble in water (1 mg/ml). Stable for at least one week at 2 to 8 °C and 6 months frozen in aliquots at -15 to -25 °C.
Analysis Note
Leupeptin gives multiple peaks on HPLC due to equilibria among three forms in solution. Purity determined using three main peaks. Majority of contaminating peptide is racemized leupeptin.Other NotesFor life science research only. Not for use in diagnostic procedures.ПараметрыQuality Level 100, biological source synthetic assay 96.5% form powder mol wt Mr 475.6 packaging pkg of 100 mg (11529048001), pkg of 25 mg (11017128001), pkg of 5 mg (11017101001), pkg of 50 mg (11034626001) Торговая марка Roche solubility H2O: 50 mg/mL shipped in wet ice storage temp. 2-8°C SMILES string OS(O)(=O)=O.CC(C)C[C@H](NC(C)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C=O.CC(C)C[C@H](NC(C)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C=O InChI 1S/2C20H38N6O4.H2O4S/c2*1-12(2)9-16(24-14(5)28)19(30)26-17(10-13(3)4)18(29)25-15(11-27)7-6-8-23-20(21)22;1-5(2,3)4/h2*11-13,15-17H,6-10H2,1-5H3,(H,24,28)(H,25,29)(H,26,30)(H4,21,22,23);(H2,1,2,3,4)/t2*15-,16-,17-;/m00./s1 InChI key CIPMKIHUGVGQTG-VFFZMTJFSA-N
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Liberase™ DH Research Grade
Кат. номер 5401089001 5401054001 General descriptionThe most commonly used method for decellularization procedures, such as tissue dissociation and cell harvesting, is based on the use of proteolytic enzymes. Proteases disrupt the extracellular matrix of tissue to allow the release of individual cells or the harvesting of cultured cells for transfer. The goal of cell isolation is to maximize the yield of dissociated cells that are viable and functionally active.
Liberase enzyme technology comprises methods for purifying clostridial collagenase isoforms to high specific activity, and for blending them together with high-specific-activity neutral protease in optimal ratios for effective dissociation of primary tissues and cultured cells. Liberase DH Research Grade contains highly purified Collagenase I and Collagenase II. These two collagenase isoforms are blended in a precise ratio to each other, and with a high concentration of Dispase®, a non-clostridial neutral protease.
Liberase enzymes are designed to increase the quality and reproducibility of tissue dissociation, and improve the viability and functionality of isolated cells. Liberase Purified Enzyme Blends replace traditional collagenase, which is a crude and variable fermentation by-product of Clostridium histolyticum.ApplicationLiberase™ DH (Dispase High) Research Grade is used for the dissociation of a broad range of tissue types, where high purity of the enzyme blend is necessary for high cell yield and viability. Bacterial by-products, such as endotoxins, are reduced up to several thousand-fold.
It has been used for the digestion of metastatic tumor fragments. It has been used for the enzymatic detachment of human embryonic stem cells colonies.
Features and Benefits- Maximize viability and yield of isolated cells
- Count on higher specific activity of the enzyme blend
- Obtain higher experimental reproducibility
Preparation Note
Stabilizers: Calcium
Working concentration: Liberase Research Grade Enzyme Working Concentration
Liberase enzymes have significantly higher specific activities than traditional collagenases. This means that the working concentration of Liberase Research Grade Purified Enzymes, expressed in mg/ml, will be lower than that of traditional collagenase.
When the application is on the Roche list of applications at www.collagenase.com, use the Liberase Research Grade concentration recommended for that application.
When the application is not included on this list, first use Liberase TM Research Grade at a concentration of 0.08–0.28 Wnsch units/ml.
The goal is to determine the best starting concentration of Liberase Research Grade Enzyme Blends. This is a starting point, and the final concentration may vary due to differences in procedure and lot-to-lot differences in traditional collagenase.
Collagenase Working Concentration
Multiply your previous collagenase working concentration (mg/ml) by its specific activity (Wnsch units/mg, [as determined above]), to obtain Wnsch units/ml. To determine how much Liberase Research Grade Enzyme Blend to use, first multiply your collagenase working concentration (in Wnsch units/ml) times the total volume of your working enzyme solution to obtain the total collagenase activity needed (Wnsch units). Divide the total collagenase activity required by the Liberase Research Grade stock concentration ( “Reconstitution and Storage”). This indicates how many milliliters of Liberase Research Grade Enzyme Blend stock solution to use in your working enzyme solution.
Storage conditions (working solution): Store unused stock solution in single-use aliquots at -15 to -25 °C. Note: Avoid repeated freezing and thawing!
Reconstitution
Reconstitute the lyophilized enzyme with injection-quality sterile water or tissue-dissociation buffer. Do not add serum or other components (e.g., albumin or protease inhibitors) to the dissociation buffer. Enzyme stability is reduced at higher concentrations and warmer temperatures (4 °C). Avoid both the above conditions.
Reconstitute the entire vial. Do not weigh individual aliquots of the lyophilizate. The introduction of moisture into the vial results in a decline in enzymatic activity.
Place vial on ice to rehydrate the lyophilized enzyme.
Gently agitate the vial at 2 to 8 °C until the enzyme is completely dissolved (max. 30 min).
Depending on the type of tissue-dissociation buffer used to dissolve Liberase Research Grade Purified Enzyme Blends, slight precipitations may be observed which readily dissolve in the diluted working solution, and have no influence on enzyme activity.
Remove an aliquot of the stock solution to prepare the working solution.
Reconstitution volume
2 ml (1 vial with 5 mg–10 mg pack size)
10 ml (1 vial with 50 mg–100 mg pack size)
Collagenase Wnsch (units/ml)
13 (1 vial with 5 mg–10 mg pack size)
26 (1 vial with 50 mg–100 mg pack size)
Total Collagenase concentration [mg/ml]
2.5 (1 vial with 5 mg–10 mg pack size)
5.0 (1 vial with 50 mg–100 mg pack size)Other Notes- This product is intended for life science research and in vitro use only. These products are not to be used for diagnostic or clinical applications, such as human islet transplantation.
- All 1st generation Liberase and Liberase Blendzyme products (Liberase HI, CI, RI, PI, Liberase Blendzyme 1, 2, 3, 4) have been replaced by the 2nd generation Liberase Research Grade and Liberase GMP grade enzyme blend portfolios. For instructions concerning the transition from previously used Liberase Enzymes
For life science research only. Not for use in diagnostic procedures.Legal InformationDispase is a registered trademark of Godo Shusei Co., Ltd.
Liberase is a trademark of RocheПараметрыQuality Level 100, form lyophilized packaging pkg of 10 mg (05401054001 [2 x 5 mg]), pkg of 100 mg (05401089001 [2 x 50 mg]) Торговая марка Roche parameter 35-37 °C optimum reaction temp. optimum pH 7.4 shipped in dry ice storage temp. 20°C
Safety InformationRIDADR NONH for all modes of transport -
Liberase™ DL Research Grade
Кат. номер 5466202001 5401160001 General descriptionLiberase™ DH Research Grade contains highly purified Collagenase I and Collagenase II. These two collagenase isoforms are blended in a precise ratio to each other, and with a high concentration of Dispase®, a non-clostridial neutral protease. Liberase™ DL (Dispase Low) Research Grade is used for the dissociation of a broad range of tissue types, where high purity of the enzyme blend is necessary for high cell yield and viability. Bacterial by-products, such as endotoxins, are reduced up to several thousand-fold.ApplicationLiberase® DL (Dispase Low) Research Grade has been used for the isolation of skin stem cells from mice. It is also used for the dissociation of a broad range of tissue types, where high purity of the enzyme blend is necessary for high cell yield and viability. Bacterial by-products, such as endotoxins, are reduced up to several thousand-fold.
Features and Benefits- Maximize viability and yield of isolated cells with an enzyme blend that has less clostripain and trypsin activity, as well as reduced endotoxin content.
- Count on higher specific activity of the enzyme blend as a result of higher Collagenase I + II purity (determined by HPLC analysis).
- Obtain higher experimental reproducibility due to higher lot-to-lot consistency.
Preparation Note
Stabilizers: Calcium
Working concentration: Liberase Research Grade Enzyme Working Concentration
Liberase enzymes have significantly higher specific activities than traditional collagenases. This means that the working concentration of Liberase Research Grade Purified Enzymes, expressed in mg/ml, will be lower than that of traditional collagenase.
When the application is on the Roche list of applications at www.collagenase.com, use the Liberase Research Grade concentration recommended for that application.
When the application is not included on this list, first use Liberase TM Research Grade at a concentration of 0.08–0.28 Wnsch units/ml.
The goal is to determine the best starting concentration of Liberase Research Grade Enzyme Blends. This is a starting point, and the final concentration may vary due to differences in procedure and lot-to-lot differences in traditional collagenase.
Collagenase Working Concentration
Multiply your previous collagenase working concentration (mg/ml) by its specific activity (Wnsch units/mg, [as determined above]), to obtain Wnsch units/ml. To determine how much Liberase Research Grade Enzyme Blend to use, first multiply your collagenase working concentration (in Wnsch units/ml) times the total volume of your working enzyme solution to obtain the total collagenase activity needed (Wnsch units). Divide the total collagenase activity required by the Liberase Research Grade stock concentration ( “Reconstitution and Storage”). This indicates how many milliliters of Liberase Research Grade Enzyme Blend stock solution to use in your working enzyme solution.
Storage conditions (working solution): Store unused stock solution in single-use aliquots at -15 to -25 °C. Note: Avoid repeated freezing and thawing!
Reconstitution
Reconstitute the lyophilized enzyme with injection-quality sterile water or tissue-dissociation buffer. Do not add serum or other components that may influence enzyme activity, such as albumin or protease inhibitors, to the dissociation buffer. Enzyme stability is reduced at higher concentrations and warmer temperatures (4 °C). Avoid both the above conditions.
Reconstitute the entire vial. Do not weigh individual aliquots of the lyophilizate. The introduction of moisture into the vial results in a decline in enzymatic activity.
Place vial on ice to rehydrate the lyophilized enzyme.
Gently agitate the vial at 2 to 8 °C until enzyme is completely dissolved (max. 30 min).
Depending on the type of tissue-dissociation buffer used to dissolve Liberase Research Grade Purified Enzyme Blends, slight precipitations may be observed readily dissolve in the diluted working solution and have no influence on enzyme activity.
Remove an aliquot of the stock solution to prepare the working solution.
Reconstitution volume
2 ml (1 vial with 5 mg–10 mg pack size)
10 ml (1 vial with 50 mg–100 mg pack size)
Collagenase Wnsch (units/ml)
13 (1 vial with 5 mg–10 mg pack size)
26 (1 vial with 50 mg–100 mg pack size)
Total Collagenase concentration [mg/ml]
2.5 (1 vial with 5 mg–10 mg pack size)
5.0 (1 vial with 50 mg–100 mg pack size)Other NotesFor life science research only. Not for use in diagnostic procedures.Legal InformationDispase is a registered trademark of Godo Shusei Co., Ltd.
Liberase is a trademark of Roche
Roche is a registered trademark of Roche Molecular Systems, Inc.ПараметрыQuality Level 100, form lyophilized packaging pkg of 100 mg (05466202001 [2 x 50 mg]), pkg of 10 mg (05401160001 [2 x 5 mg]) Торговая марка Roche parameter 35-37 °C optimum reaction temp. optimum pH 7.4 shipped in dry ice storage temp. 20°C
Safety InformationRIDADR NONH for all modes of transport -
Liberase™ TH Research Grade
Кат. номер 5401151001 5401135001 General descriptionLiberase TH Research Grade contains highly purified Collagenase I and Collagenase II. These two collagenase isoforms are blended in a precise ratio to each other and with a high concentration of Thermolysin, a non-clostridial neutral protease.ApplicationLiberase™ TH (Thermolysin High) Research Grade is used for the dissociation of a broad range of tissue types, where high purity of the enzyme blend is necessary for high cell yield and viability. Bacterial by-products, such as endotoxins, are reduced up to several thousand-fold.
Features and Benefits- Maximize viability and yield of isolated cells
- Count on higher specific activity of the enzyme blend
- Obtain higher experimental reproducibility
- Increase safety
Preparation Note
Stabilizers: Calcium
Working concentration: Liberase Research Grade Enzyme Working Concentration:
Liberase enzymes have significantly higher specific activities than traditional collagenases. This means that the working concentration of Liberase Research Grade Purified Enzymes, expressed in mg/ml, will be lower than that of traditional collagenase.
When the application is on the Roche list of applications at www.collagenase.com, use the Liberase Research Grade concentration recommended for that application.
When the application is not included on this list, first use Liberase TM Research Grade at a concentration of 0.08–0.28 Wnsch units/ml.
The goal is to determine the best starting concentration of Liberase Research Grade Enzyme Blends. This is a starting point, and the final concentration may vary due to differences in procedure and lot-to-lot differences in traditional collagenase.
Collagenase Working Concentration:
Multiply your previous collagenase working concentration (mg/ml) by its specific activity (Wnsch units/mg, [as determined above]), to obtain Wnsch units/ml. To determine how much Liberase Research Grade Enzyme Blend to use, first multiply your collagenase working concentration (in Wnsch units/ml) times the total volume of your working enzyme solution to obtain the total collagenase activity needed (Wnsch units). Divide the total collagenase activity required by the Liberase Research Grade stock concentration ( “Reconstitution and Storage”). This indicates how many milliliters of Liberase Research Grade Enzyme Blend stock solution to use in your working enzyme solution.
Storage conditions (working solution): Store unused stock solution in single-use aliquots at -15 to -25 °C. Note: Avoid repeated freezing and thawing!
Reconstitution
Reconstitute the lyophilized enzyme with injection-quality sterile water or tissue-dissociation buffer. Do not add serum or other components such as albumin or protease inhibitors, to the dissociation buffer. Enzyme stability is reduced at higher concentrations and warmer temperatures (4 °C). Avoid both the above conditions.
Reconstitute the entire vial. Do not weigh individual aliquots of the lyophilizate. The introduction of moisture into the vial results in a decline in enzymatic activity.
Place vial on ice to rehydrate the lyophilized enzyme.
Gently agitate the vial at 2 to 8 °C until enzyme is completely dissolved (max. 30 min).
Depending on the type of tissue-dissociation buffer used to dissolve Liberase Research Grade Purified Enzyme Blends, slight precipitations may be observed which readily dissolve in the diluted working solution and have no influence on enzyme activity.
Remove an aliquot of the stock solution to prepare the working solution.
Reconstitution volume
2 ml (1 vial with 5 mg–10 mg pack size), 10 ml (1 vial with 50 mg–100 mg pack size)
Collagenase Wnsch (units/ml)
13 (1 vial with 5 mg–10 mg pack size), 26 (1 vial with 50 mg–100 mg pack size)
Total Collagenase concentration [mg/ml]
2.5 (1 vial with 5 mg–10 mg pack size), 5.0 (1 vial with 50 mg–100 mg pack size)Other NotesFor life science research only. Not for use in diagnostic procedures.Legal InformationLiberase is a trademark of RocheПараметрыQuality Level 100, form lyophilized packaging pkg of 10 mg (05401135001 [2 x 5 mg]), pkg of 100 mg (05401151001 [2 x 50 mg]) Торговая марка Roche parameter 35-37 °C optimum reaction temp. optimum pH 7.4 shipped in dry ice storage temp. 20°C
Safety InformationRIDADR NONH for all modes of transport -
5401020001
Liberase™ TL Research Grade 5401020001
General descriptionThe most commonly used method for decellularization procedures, such as tissue dissociation and cell harvesting, is based on the use of proteolytic enzymes. Proteases disrupt the extracellular matrix of tissue to allow the release of individual cells or the harvesting of cultured cells for transfer. The goal of cell isolation is to maximize the yield of dissociated cells that are viable and functionally active.
Liberase Purified Enzyme Blends
Liberase enzymes are blends of highly purified enzymes, designed to increase the quality and reproducibility of tissue dissociation, and improve the viability and functionality of isolated cells. Liberase Purified Enzyme Blends replace traditional collagenase, which is a crude and variable fermentation by-product of Clostridium histolyticum.
Liberase Enzyme Technology
Liberase enzyme technology comprises methods for purifying clostridial collagenase isoforms to high specific activity, and for blending them together with high-specific-activity neutral protease in optimal ratios for effective dissociation of primary tissues and cultured cells.
For additional information, such as manufacturing, technology, or storage, see the Liberase Research Grade Insightsdocument. Further reading on the topic can also be found in the Documents tab.
Roche provides special-quality preparations of Liberase Purified Enzyme Blends, manufactured to meet GMP requirements. For more information on the GMP-grade preparations.
Important Notes:- This product is intended for life science research and in vitro use only. These products are not to be used for diagnostic or clinical applications, such as human islet transplantation.
- All 1st generation Liberase and Liberase Blendzyme products (Liberase HI, CI, RI, PI, Liberase Blendzyme 1, 2, 3, 4) have been replaced by the 2nd generation Liberase Research Grade and Liberase GMP grade enzyme blend portfolios. For instructions concerning the transition from previously used Liberase Enzymes.
ApplicationLiberase™ TL (Thermolysin Low) Research Grade is used for the dissociation of a broad range of tissue types, where high purity of the enzyme blend is necessary for high cell yield and viability. Bacterial by-products, such as endotoxins, are reduced up to several thousand-fold.
This product has been used for the isolation of pancreatic islets from mice.Features and BenefitsLiberase TL Research Grade contains highly purified Collagenase I and Collagenase II. These two collagenase isoforms are blended in a precise ratio to each other and with a low concentration of Thermolysin, a non-clostridial neutral protease.- Maximize viability and yield of isolated cells with an enzyme blend that has less clostripain and trypsin activity, as well as reduced endotoxin content.
- Count on higher specific activity of the enzyme blend as a result of higher Collagenase I + II purit(determined by HPLC analysis)
- Obtain higher experimental reproducibility due to higher lot-to-lot consistency.
- Increase safety with an enzyme that is free of any mammalian or avian tissue-derived raw materials.
Preparation NoteStabilizers: Calcium
Working concentration: Liberase Research Grade Enzyme Working Concentration
Liberase enzymes have significantly higher specific activities than traditional collagenases. This means that the working concentration of Liberase Research Grade Purified Enzymes, expressed in mg/ml, will be lower than that of traditional collagenase.
When the application is on the Roche list of applications at www.collagenase.com, use the Liberase Research Grade concentration recommended for that application.
When the application is not included on this list, first use Liberase TM Research Grade at a concentration of 0.08–0.28 Wnsch units/ml.
The goal is to determine the best starting concentration of Liberase Research Grade Enzyme Blends. This is a starting point, and the final concentration may vary due to differences in procedure and lot-to-lot differences in traditional collagenase.
Collagenase Working Concentration
Multiply your previous collagenase working concentration (mg/ml) by its specific activity (Wnsch units/mg, as determined above), to obtain Wnsch units/ml. To determine how much Liberase Research Grade Enzyme Blend to use, first multiply your collagenase working concentration (in Wnsch units/ml) times the total volume of your working enzyme solution to obtain the total collagenase activity needed (Wnsch units). Divide the total collagenase activity required by the Liberase Research Grade stock concentration ( “Reconstitution and Storage”). This indicates how many milliliters of Liberase Research Grade Enzyme Blend stock solution to use in your working enzyme solution.
Storage conditions (working solution): Store unused stock solution in single-use aliquots at -15 to -25 °C. Note: Avoid repeated freezing and thawing!ReconstitutionReconstitute the lyophilized enzyme with injection-quality sterile water or tissue-dissociation buffer. Do not add serum or other components that may influence enzyme activity, such as albumin or protease inhibitors, to the dissociation buffer. Enzyme stability is reduced at higher concentrations and warmer temperatures (4 °C). Avoid both the above conditions.
Reconstitute the entire vial. Do not weigh individual aliquots of the lyophilizate. The introduction of moisture into the vial results in a decline in enzymatic activity.
Place vial on ice to rehydrate the lyophilized enzyme.
Gently agitate the vial at 2 to 8 °C until enzyme is completely dissolved (max. 30 min).
Depending on the type of tissue-dissociation buffer used to dissolve Liberase Research Grade Purified Enzyme Blends, slight precipitations may be observed which readily dissolve in the diluted working solution and have no influence on enzyme activity.
Remove an aliquot of the stock solution to prepare the working solution
Reconstitution volume
2 ml (1 vial with 5 mg–10 mg pack size), 10 ml (1 vial with 50 mg–100 mg pack size)
Collagenase Wnsch (units/ml)
13 (1 vial with 5 mg–10 mg pack size), 26 (1 vial with 50 mg–100 mg pack size)
Total Collagenase concentration mg/ml
2.5 (1 vial with 5 mg–10 mg pack size), 5.0 (1 vial with 50 mg–100 mg pack size)Storage and StabilityStore at -15–-25 °C. (Store dry. Please note that the expiration date is not printed on the individual vials.)Other NotesFor life science research only. Not for use in diagnostic procedures.Legal InformationLiberase is a trademark of RocheПараметрыQuality Level 100 form lyophilized packaging pkg of 10 mg (2x5 mg) mfr. no. Roche parameter 35-37 °C optimum reaction temp. optimum pH 7.4 shipped in dry ice
Safety Information