Roche® Life Science Products
- Сортировать:
- Вид таблицей
-
Cell Death Detection ELISA 11544675001
General descriptionThe Cell Death Detection ELISA serves as photometric enzyme immunoassay for the qualitative and quantitative in vitro determination of cytoplasmic histone-associated DNA fragments (mono- and oligonucleosomes) after induced cell death. This assay is based on the quantitative sandwich-enzyme immunoassay-principle using mouse monoclonal antibodies directed against DNA and histones, respectively. The antibodies used are not species specific.
Specificity
Anti-histone antibody reacts with the histones H1, H2A, H2B, H3, and H4 of various species (e.g., human, mouse, rat, hamster, cow, opossum, and Xenopus). Anti-DNA-POD antibody binds to ss- and dsDNA. Therefore, the ELISA allows the detection of mono- and oligonucleosomes from various species, and may be applied to measure apoptotic cell death in many different cell systems.ApplicationCell Death Detection ELISA has been used to measure Caspase-3 activity, intracellular reactive oxygen species (ROS) and cell apoptosis.
For research use only. Not for use in diagnostic procedures.
Specific determination of mono- and oligonucleosomes in the cytoplasmic fraction of cell lysates.
Packaging
1 kit containing 9 components.
Preparation Note
Working solution: Coating solution
Predilute 1 ml Coating buffer conc. with 9 ml double dist. water. Shortly before use, dilute 1 ml anti-histone antibody (reconstituted) with 9 ml Coating buffer.
Washing solution
Warm the Washing buffer concentrate to 15 to 25 °C and dilute 40 ml in 360 ml double distilled water. Mix thoroughly.
Sample solution
Preparation of the sample solution depends on the cell system used and the extent of cell death.
Example: Dilute 25 µl of sample in 225 µl Incubation buffer.
Conjugate solution
Dilute 1 ml Anti-DNA-POD (reconstituted) with 9 ml Incubation buffer.
Substrate solution
Depending on the number of samples tested, dissolve 1, 2, or 3 tablets in 5, 10, or 15 ml Substrate buffer.
Allow to come to 15 to 25 °C before use.
Note: The ABTS solution is light sensitive over extended periods of time.
Storage conditions (working solution): Anti-histone
2 to 8 °C for 2 months
Anti-DNA-POD
2 to 8 °C for 2 months
Coating solution
Prepare immediately before use!
Washing solution
2 to 8 °C for 2 months
Sample solution
2 to 8 °C for 2 months
Conjugate solution
Prepare immediately before use!
Substrate solution
2 to 8 °C for 1 month, protected from light.
Reconstitution
Anti-histone
Reconstitute the lyophilizate in 1 ml double-distilled water for 10 minutes.Mix thoroughly.
Anti-DNA-POD
Reconstitute the lyophilizate in 1 ml double-distilled water for 10 minutes. Mix thoroughly.Other NotesFor life science research only. Not for use in diagnostic procedures.Параметрыusage sufficient for 96 tests Quality Level 100, Торговая марка Roche shipped in wet ice storage temp. 2-8°C
Safety Informationpictograms GHS07 signalword Warning hcodes H317 - H319 pcodes P261 - P280 - P333 + P313 - P337 + P313 - P362 + P364 - P501 RIDADR NONH for all modes of transport WGK Germany WGK 2 Flash Point F does not flash Flash Point C does not flash Узнать цену -
Cell Proliferation ELISA, BrdU (chemiluminescent) 11669915001
General descriptionChemiluminescent immunoassay for the quantification of cell proliferation, based on the measurement of BrdU incorporation during DNA synthesis: a nonradioactive alternative to the [3H]-thymidine incorporation assay.
Specificity
The antibody conjugate reacts with the thymidine analogue 5-bromo-2′-deoxyuridine (BrdU) and with BrdU incorporated into DNA. For binding to BrdU incorporated into the DNA, the BrdU-labeled DNA has to be denatured. The antibody does not cross-react with any endogenous cellular components such as thymidine, uridine, or DNA.ApplicationCell Proliferation ELISA, BrdU (chemiluminescent) has been used:- for bromodeoxyuridine (BrdU) proliferation assay in ReN and primary astrocytes
- to test the effect on imatinib on the proliferation of neuroblastoma cells using BrdU incorporation assay
- to check the effect of palbociclib on cell cycle inhibition using BrdU incorporation in pancreatic patient-derived primary cell lines
The Cell Proliferation ELISA, BrdU (chemiluminescent) belongs to the second, improved generation of kits for measuring DNA synthesis. It is a precise, fast, and simple nonradioactive alternative to quantitate cell proliferation based on the measurement of BrdU incorporation during DNA synthesis in replicating (cycling) cells. Thus, the Cell Proliferation ELISA can be used in many different in vitro cell systems. For example:- Detection and quantification of cell proliferation induced by growth factors and cytokines
- Determination of the inhibitory or stimulatory effects of various compounds on cell proliferation in environmental and biomedical research, and in the food, cosmetic, and pharmaceutical industries
- Measurement of the immunoreactivity of lymphocytes, stimulated by mitogens or antigens
- Analysis of the chemosensitivity of tumor cells to different cytostatic drugs in medical research
Packaging
1 kit containing 7 components.Other NotesFor life science research only. Not for use in diagnostic procedures.Параметрыusage sufficient for ≤1,000 tests Quality Level 100, Торговая марка Roche shipped in wet ice storage temp. 2-8°C
Safety Informationpictograms GHS02,GHS07 signalword Danger hcodes H225 - H317 - H319 pcodes P210 - P233 - P261 - P280 - P303 + P361 + P353 - P370 + P378 RIDADR UN1170 - class 3 - PG 2 - Ethanol, solution WGK Germany WGK 2 Узнать цену -
Cell Proliferation ELISA, BrdU (colorimetric) 11647229001
General descriptionColorimetric immunoassay for the quantification of cell proliferation, based on the measurement of BrdU incorporation during DNA synthesis: a nonradioactive alternative to the [3H]-thymidine incorporation assay.
Specificity
The antibody conjugate reacts with the thymidine analogue 5-bromo-2′-deoxyuridine (BrdU) and with BrdU incorporated into DNA. For binding to BrdU incorporated into the DNA, the BrdU-labeled DNA has to be denatured. The antibody does not cross-react with any endogenous cellular components such as thymidine, uridine, or DNA.ApplicationFor research use only. Not for use in diagnostic procedures.
The Cell Proliferation ELISA, BrdU (colorimetric) belongs to the second, improved generation of kits for measuring DNA synthesis. It is a precise, fast, and simple colorimetric alternative to quantitate cell proliferation based on the measurement of BrdU incorporation during DNA synthesis in replicating (cycling) cells. Thus, the Cell Proliferation ELISA can be used in many different in vitro cell systems. For example:- Detection and quantification of cell proliferation induced by growth factors and cytokines
- Determination of the inhibitory or stimulatory effects of various compounds on cell proliferation in environmental and biomedical research, and in the food, cosmetic, and pharmaceutical industries
- Measurement of the immunoreactivity of lymphocytes, stimulated by mitogens or antigens
- Analysis of the chemosensitivity of tumor cells to different cytostatic drugs in medical research
- Testing of biocompatibility of various scaffolds, employed in bone tissue engineering, for bone cell growth
Packaging
1 kit containing 6 components.
Preparation Note
Working concentration: The kit antibody (Anti-BrdU-peroxidase) has, after reconstitution, a concentration of 7.5 U/ml, after dilution 0.075 U/ml. Instead of the kit antibody, you can also use the Anti-BrdU-Antibody, Fab fragments.This antibody is double-concentrated, so you have to dilute it after reconstitution in 1 ml with 1:200.
Working solution: BrdU labeling solution
Dilute BrdU labeling reagent 1:100 with sterile culture medium (resulting concentration: 100 µM BrdU).
For one 96-well MP, 1 ml BrdU labeling solution is required if the cells were cultured in 100 µl /well (10 µl/well) and 2 ml BrdU labeling solution is required if the cells were cultured in 200 µl/well (20 µl/well).
Anti-BrdU-peroxidase stock solution
Dissolve Anti-BrdU-peroxidase in 1.1 ml double-dist. water for 10 minutes and mix thoroughly.
Anti-BrdU-peroxidase working solution
Dilute Anti-BrdU-peroxidase stock solution 1:100 with antibody dilution solution. For one 96-well MP dilute 100 µl Anti-BrdU-peroxidase stock solution in 10 ml antibody dilution solution
Washing solution
Dilute Washing buffer concentrate 1:10 with double-dist. water.
For one 96-well MP dilute 10 ml Washing buffer concentrate with 90 ml double-dist. water.
Storage conditions (working solution): BrdU labeling solution
The undiluted BrdU labeling reagent (1000x): At 2 to 8 °C for several months protected from light.
The diluted BrdU labeling reagent: At 2 to 8 °C stable for several weeks. Store protected from light. For long-term storage it is recommended to store the BrdU labeling solution in aliquots at -15 to -25 °C.
Anti-BrdU-peroxidase stock solution
At 2 to 8 °C for several months. For long-term storage it is recommended to store the solution in aliquots at -15 to -25 °C.
Anti-BrdU-peroxidase working solution
Prepare shortly before use. Do not store.
Washing solution
At 2 to 8 °C for several weeks.
Reconstitution
The working solution for the antibody should be phosphate buffered saline containing 1% BSA, pH 7.4Other NotesFor life science research only. Not for use in diagnostic procedures.Параметрыusage sufficient for ≤1,000 tests Quality Level 100, Торговая марка Roche shipped in wet ice storage temp. 2-8°C
Safety Informationpictograms GHS02,GHS07 signalword Danger hcodes H225 - H317 - H319 pcodes P210 - P233 - P261 - P280 - P303 + P361 + P353 - P370 + P378 RIDADR UN1170 - class 3 - PG 2 - Ethanol, solution WGK Germany WGK 2 Узнать цену -
Cell Proliferation Kit I (MTT) 11465007001
General descriptionThe Cell Proliferation Kit I (MTT) is a colorimetric assay for the nonradioactive quantification of cellular proliferation, viability, and cytotoxicity. Sample material is either adherent or suspension cells cultured in 96-well microplates.
Colorimetric assays analyze the number of viable cells by the cleavage of tetrazolium salts added to the culture medium. This technique requires neither washing nor harvesting of cells, and the complete assay, from microculture to data analysis by an ELISA reader, is performed in the same microplate.
MTT was the first tetrazolium salt described. It is cleaved to formazan by enzymes of the endoplasmic reticulum. This bioreduction occurs in viable cells only, and is related to NAD(P)H production through glycolysis. Therefore, the amount of formazan dye formed directly correlates to the number of metabolically active cells in the culture.
Colorimetric assay (MTT based) for the nonradioactive quantification of cellular proliferation, viability, and cytotoxicity.ApplicationThe Cell Proliferation Kit I (MTT) is used for the nonradioactive, spectrophotometric quantification of cell proliferation and viability in cell populations using the 96-well-plate format. It can be used for:- Measurement of cell proliferation in response to growth factors, cytokines, mitogens, and nutrients
- Analysis of cytotoxic and cytostatic compounds, such as anti-cancer drugs and other pharmaceutical compounds
- Assessment of growth-inhibitory antibodies and physiological mediators
- Testing of biocompatibility of various scaffolds, employed in bone tissue engineering, for bone cell growth
Features and Benefits- Safe and easy: Eliminate radioactive isotopes, washing steps, and additional reagents.
- Accurate: The absorbance obtained strongly correlates to the cell number.
- Sensitive: Detect low cell numbers.
- Fast: Process a large number of samples using a multi-well ELISA reader.
The assay is based on the cleavage of the tetrazolium salt MTT in the presence of an electron-coupling reagent. The water-insoluble formazan salt produced must be solubilized in an additional step. Cells grown in a 96-well tissue culture plate are incubated with the MTT solution for approximately 4 hours. After this incubation period, a water-insoluble formazan dye is formed. After solubilization, the formazan dye is quantitated using a scanning multi-well spectrophotometer (ELISA reader). The measured absorbance directly correlates to the number of viable cells.
Cell proliferation and viability assays are of particular importance for routine applications in cell biology. Tetrazolium salts (e.g., MTT, XTT, WST-1) are particularly useful for this type of analysis. Tetrazolium salts are cleaved to formazan by the succinate-tetrazolium reductase system (EC 1.3.99.1) which belongs to the respiratory chain of the mitochondria, and is only active in metabolically intact cells (see below).
Packaging
1 kit containing 2 components.
Preparation Note
Storage conditions (working solution): 2 to 8 °C
After thawing, the MTT labeling reagents may be stored protected from light at 2 to 8 °C for up to 4 weeks, in which case reagent filtration through a 0.2 µm pore size membrane is recommended.Other NotesFor life science research only. Not for use in diagnostic procedures.Параметрыform liquid Quality Level 100, usage sufficient for 2,500 tests packaging pkg of 1 kit Торговая марка Roche storage condition protect from light application(s) cell analysis: suitable, detection: suitable, tissue culture: suitable max 550-600 nm detection method colorimetric shipped in dry ice storage temp. 20°C
Safety Informationpictograms GHS05 signalword Danger hcodes H318 pcodes P280 - P305 + P351 + P338 + P310 RIDADR NONH for all modes of transport WGK Germany WGK 2 Flash Point F does not flash Flash Point C does not flash Узнать цену -
Cell Proliferation Kit II (XTT) 11465015001
General descriptionThe Cell Proliferation Kit II (XTT) is a colorimetric assay for the nonradioactive quantification of cellular proliferation, viability, and cytotoxicity. Sample material is either adherent or suspension cells cultured in 96-well microplates.
Colorimetric assays analyze the number of viable cells by the cleavage of tetrazolium salts added to the culture medium. This technique requires neither washing nor harvesting of cells, and the complete assay, from microculture to data analysis by an ELISA reader, is performed in the same microplate.
More recently, the tetrazolium salt XTT was described. In contrast to MTT, the cleavage product of XTT is soluble in water; therefore, a solubilization step is not required. The tetrazolium salt XTT is cleaved to formazan by a complex cellular mechanism. This bioreduction occurs in viable cells only, and is related to NAD(P)H production through glycolysis. Therefore, the amount of formazan dye formed directly correlates to the number of metabolically active cells in the culture.ApplicationThe Cell Proliferation Kit II (XTT) is used for the nonradioactive, spectrophotometric quantification of cell proliferation and viability in cell populations using the 96-well-plate format. It can be used for:- Measurement of cell proliferation in response to growth factors, cytokines, mitogens, and nutrients.
- Analysis of cytotoxic and cytostatic compounds, such as anti-cancer drugs and other pharmaceutical compounds.
- Assessment of growth-inhibitory antibodies and physiological mediators that inhibit cell growth.
- Testing of biocompatibility of various scaffolds, employed in bone tissue engineering, for bone cell growth.
- cell viability assay.
Features and Benefits- Safe and easy: Eliminate radioactive isotopes, washing steps, and additional reagents.
- Accurate: The absorbance obtained strongly correlates to the cell number.
- Sensitive: Detect low cell numbers.
- Fast: Process a large number of samples using a multi-well ELISA reader.
Packaging
1 kit containing 2 components.
Principle
The assay is based on the cleavage of the tetrazolium salt XTT in the presence of an electron-coupling reagent, producing a soluble formazan salt. This conversion only occurs in viable cells. Cells grown in a 96-well tissue culture plate are incubated with the XTT labeling mixture for 2 - 20 hours. After this incubation period, the formazan dye formed is quantitated using a scanning multi-well spectrophotometer (ELISA reader). The measured absorbance directly correlates to the number of viable cells.
Cell proliferation and viability assays are of particular importance for routine applications in cell biology. Tetrazolium salts (e.g., MTT, XTT, WST-1) are particularly useful for this type of analysis. Tetrazolium salts are cleaved to formazan by the succinate-tetrazolium reductase system (EC 1.3.99.1) which belongs to the respiratory chain of the mitochondria, and is only active in metabolically intact cells.
Preparation Note
Working solution: Preparation of solutions
Thaw XTT labeling reagent and electron-coupling reagent, respectively in a water bath at 37 °C. Mix each vial thoroughly to obtain a clear solution.
XTT labeling mixture
To perform a cell proliferation assay (XTT) with one microplate (96 wells) mix 5 ml XTT labeling reagent with 0.1 ml electron coupling reagent.
Note: To obtain reliable results thaw and mix XTT labeling reagent and electron coupling reagent immediately before use.
Working instruction
Cells are grown in microplates (tissue culture grade, 96 wells, flat bottom) in a final volume of 100 µl culture medium per well, according to the media needs of the cells in a humidified atmosphere (e.g., 37 °C, 6.5% CO2).
The incubation period of the cell cultures depends on the particular experimental approach and on the cell line, used for the assay. For most experimental setups, the incubation of cells for 24 to 96 hours is appropriate.
After the incubation period, add to each well 50 µl of the XTT labeling mixture, prepared as described above (final XTT concentration 0.3 mg/ml).
Incubate the microplate for 4 to 24 hours in a humidified atmosphere (e.g., 37 °C, 6.5% CO2).
Note: The incubation time varies with the individual experimental setup (e.g., cell type and cell concentration, used). Therefore, we recommend to measure the absorption as described at different time points after addition of XTT labeling mixture (e.g., 4, 6, 8, 12, and 18 hours) using one and the same microplate to determine the optimal incubation period for the particular experimental setup.
Storage conditions (working solution): Thaw reagents immediately before use. It is recommended to prepare appropriate aliquots [5 ml XTT labeling reagent and 0.1 ml electron coupling reagent are required for the performance of the assay with one microplate (96 wells)]
Note: Avoid repeated thawing and freezing.Other NotesFor life science research only. Not for use in diagnostic procedures.Параметрыform liquid Quality Level 100, usage sufficient for 2,500 tests packaging pkg of 1 kit Торговая марка Roche storage condition protect from light application(s) cell analysis: suitable, detection: suitable, tissue culture: suitable max 450-500 nm detection method colorimetric shipped in dry ice storage temp. 20°C
Safety InformationRIDADR NONH for all modes of transport WGK Germany nwg Flash Point F does not flash Flash Point C does not flash Узнать цену -
Cell Viability Imaging Kit 5x96 6432379001
General descriptionThe Cell Viability Imaging Kit is a simple and rapid way to measure mammalian cell viability using fluorescence microscopy and automated imaging platforms, such as the Cellavista System. Optimized for 96-well plates, the numbers of viable and dead cells, as well as total cell numbers, are determined simultaneously for the area of the well selected.
The kit is designed for 5 x 96 reactions for use in medium to high throughput analysis of cell behavior.- Determine total cell number using fluorescence staining of cell nuclei.
- Fluorimetrically determine the percentage of dead cells based on membrane permeability.
- Verify the labeling of viable cells quantified by fluorescence staining using a vital dye
ApplicationCell quantification and cell viability control are very important methods in cell biology. This includes both life science research and industrial applications. The simple, accurate and fast “one step assay” makes the Cell Viability Imaging Kit an ideal tool for quality control of cells in cellular workflows, especially for medium to high throughput applications.
Features and Benefits
Cell counting and cell viability are essential for cellular workflows. The Roche Cell Viability Imaging Kit meets the following requirements:- No risk of losing cells in the one step assay: There are no fixation or washing steps.
- Save time and resources: Total assay time for a 1 x 96 well microplate is only 35 minutes.
- Obtain highly reproducible results using automated imaging and data evaluation with Roches Cellavista System.
Packaging
1 kit containing 3 components.
Preparation Note
Storage conditions (working solution): Vial 3: The product is stable until the expiration date printed on the label when stored at -15 to -25 °C. Contents can be frozen and thawed at least 5 times after preparation of solution.
Storage and Stability
Store at -15–-25 °C. (Vial 1 and Vial 2 can be frozen and thawed at least 5 times. Vial 3 can be frozen and thawed at least 5 times after preparation of solution.)Other NotesFor life science research only. Not for use in diagnostic procedures.ПараметрыQuality Level 100, usage sufficient for 5 96-well plate(s) packaging pkg of 1 kit Торговая марка Roche storage condition protect from light fluorescence ex 361 nm; em 486 nm (nuclei dye), ex 490 nm; em 515 nm (viable dye), ex 535 nm; em 617 nm (dead dye) detection method fluorometric shipped in dry ice storage temp. 20°C (15°C to 25°C)
Safety Informationpictograms GHS08 signalword Warning hcodes H341 pcodes P201 - P202 - P280 - P308 + P313 - P405 - P501 RIDADR NONH for all modes of transport WGK Germany WGK 1 Flash Point F Not applicable Flash Point C Not applicable Узнать цену -
Cellular DNA Fragmentation ELISA 11585045001
General descriptionThe Cellular DNA Fragmentation ELISA is a photometric enzyme-linked immunosorbent assay (ELISA) for the detection of 5-bromo-2-deoxy-uridine (BrdU)-labeled DNA fragments in culture supernatants and cell lysates. BrdU is used as a metabolic labeling agent by the nuclear DNA of target cells. This BrdU-labeled DNA can be detected easily and quantified using a monoclonal antibody against BrdU in an ELISA. This kit is a non-radioactive alternative to the [3H]-thymidine release assay, the [51Cr]-release assay, and the [3H]-thymidine DNA fragmentation assay.ApplicationFor research use only. Not for use in diagnostic procedures.
The Cellular DNA Fragmentation ELISA is for the quantification of BrdU-labeled DNA fragments either released from cells during necrosis or cell-mediated cytotoxicity, or within the cytoplasm of apoptotic cells.
Features and Benefits- Nonradioactive assay system
- Sensitive (1 x 103 cells/well)
- Low assay background
- No additional dilution steps required
- No washing of the cells required
- Results obtained correlate to those obtained by standard methods
- Denaturation/fixation of DNA in the microplate by microwave irradiation
- Fast (4.5 - 5.5 hours)
Packaging
1 kit containing 8 components.
Preparation Note
Working solution: Solubility: soluble in water, aqueous buffers, and dimethylformamide. Can be dissolved in methanol. A clear solution is achieved at 20 mg/ml.
Storage conditions (working solution): Storage of coated plates: 1 week at 2 to 8 °C.
The anti-BrdU-POD stock solution is stable at 2 to 8 °C for several months; for long- term storage it is recommended to store the solution in aliquots at -15 to -25 °C;
The anti-BrdU-working solution should not be stored.
For the anti-DNA-clone MCA-33 we do no have stability data. However it should be no problem to aliquot and store this non-conjugated antibody at -15 to -25 °C for longer storage.Other NotesFor life science research only. Not for use in diagnostic procedures.Параметрыusage sufficient for ≤500 tests Quality Level 100, Торговая марка Roche shipped in wet ice storage temp. 2-8°C
Safety Informationpictograms GHS07 signalword Warning hcodes H317 - H319 - H412 pcodes P261 - P273 - P280 - P333 + P313 - P337 + P313 - P362 + P364 RIDADR NONH for all modes of transport WGK Germany WGK 2 Flash Point F does not flash Flash Point C does not flash Узнать цену -
CHAPS
Empirical Formula (Hill Notation): C32H58N2O7S · xH2O CAS Number: 75621-03-3 Molecular Weight: 614.88 (anhydrous basis)
Кат. номер 10810126001 10810118001 General description3-[(3-Cholamidopropyl)dimethyl-ammonio]-1-propane sulfonateApplicationSolubilization of membrane proteins. Combines the properties of both the sulfobetaine-type and bile-acid detergents.
Physical form
Crystals
Preparation Note
Working concentration: PAGE: 6.5 to 13 mM
Storage conditions (working solution): Do not store in solution, because precipitation may occur.Other NotesFor life science research only. Not for use in diagnostic procedures.Параметрыassay 98% (from N) mol wt 614.88 g/mol (anhydrous basis) packaging pkg of 10 g (10810118001), pkg of 50 g (10810126001) Торговая марка Roche application(s) dialysis: suitable CMC 4 mM mp 157 °C (dec.) (lit.) solubility >50% (Aqueous solutions) absorption <0.06% at 278 nm in water at 1 % (w/v) (Ease of removal*: ++
Note:
* Arbitrary scale ranges from ++ to )shipped in wet ice storage temp. 2-8°C SMILES string O.C[C@H](CCC(=O)NCCC[N+](C)(C)CCCS([O-])(=O)=O)[C@H]1CC[C@H]2[C@@H]3[C@H](O)C[C@@H]4C[C@H](O)CC[C@]4(C)[C@H]3C[C@H](O)[C@]12C InChI 1S/C32H58N2O7S.H2O/c1-21(8-11-29(38)33-14-6-15-34(4,5)16-7-17-42(39,40)41)24-9-10-25-30-26(20-28(37)32(24,25)3)31(2)13-12-23(35)18-22(31)19-27(30)36;/h21-28,30,35-37H,6-20H2,1-5H3,(H-,33,38,39,40,41);1H2/t21-,22+,23-,24-,25+,26+,27-,28+,30+,31+,32-;/m1./s1 InChI key SJCUTFKCLFLIFE-JWTJKVBLSA-N
Safety InformationRIDADR NONH for all modes of transport Узнать цену -
Chromozym PK 10378445001
General descriptionChromozym® PK is a non-radioactive amidolytic chromogenic substrate mostly used in kinetic analysis of plasma kallikrein.ApplicationUse Chromozym® PK as a substrate for the determination of serine proteases, specifically plasma kallikrein in citrated plasma.
Quality
Contaminant: <0.5% free 4-nitraniline
Sequence
Benzoyl-prolyl-phenalanyl-arginine-4-nitranilide-acetate
Principle
Chromozym PK is cleaved by plasma kallikrein into a residual peptide and free 4-nitraniline, which is measured at 405 nm. The absorbance difference per minute is used for the determination of the kallikrein activity in U/ml.
Preparation Note
Activator: Mg2+ (Km = 3.0 mM, pH 7.4, 25 °C); other divalent metal ions (Mn2+, Fe2+, Zn2+)
Working concentration: Approximately 0.5 mM
Storage conditions (working solution): 2 to 8 °C
An aqueous solution of Chromozym PK (1.3 mM) is stable for at least 4 weeks if stored at 2 to 8 °C.Other NotesFor life science research only. Not for use in diagnostic procedures.Legal InformationChromozym is a registered trademark of Pentapharm AG, BaselПараметрыdescription Molecular Formula: C33H39N8O6COOCH3 Quality Level 100, assay 90% (enzymatic) form powder mol wt Mr 702.9 packaging pkg of 20 mg Торговая марка Roche shipped in ambient storage temp. room temp
Safety InformationRIDADR NONH for all modes of transport WGK Germany WGK 1 Flash Point F Not applicable Flash Point C Not applicable Узнать цену -
Chromozym PL 10378461001
General descriptionChromozym® PL is a chromogenic substrate. It is a synthetic tripeptide substrate and is more plasmin-specific.ApplicationChromozym® PL has been used:- to determine the plasmin enzymatic activity using human umbilical vein endothelial cells (HUVEC) cell extracts
- as a chromogenic substrate for proteolytic activity in murine lung homogenates
- as a substrate of plasmin in plasminogen activation assays
Quality
Purity: 90% Tosyl-Gly-Pro-Lys-4-nitranilide acetate (enzymatic)
Contaminant: <0.5% free 4-nitraniline
Formula: C26H35N6O7SCOOCH3
Sequence
Tosyl-glycyl-polyl-lysine-4-nitranilide-acetate
Physical form
Powder
Preparation Note
Working concentration: Approximately 0.3 to 0.6 mM
Storage conditions (working solution): 2 to 8 °C
A substrate solution (3 mM) is stable for at least two weeks if stored at 2 to 8 °C. Avoid contamination with germs.Other NotesFor life science research only. Not for use in diagnostic procedures.Legal InformationChromozym is a registered trademark of Pentapharm AG, BaselПараметрыassay 90% Quality Level 100, form powder packaging pkg of 20 mg Торговая марка Roche shipped in ambient storage temp. room temp
Safety InformationRIDADR NONH for all modes of transport WGK Germany WGK 2 Flash Point F Not applicable Flash Point C Not applicable Узнать цену -
Chromozym t-PA 11093037001
General descriptionN-Methylsulfonyl-D-Phe-Gly-Arg-4-nitranilide acetateApplicationUse Chromozym® t-PA (tissue plasminogen activator) (C24H32N8O7S x CH3COOH) as a substrate for the determination of t-PA, both in purified preparations and in cell culture supernatants. Moreover, it can be applied to evaluate the content of one-chain and two-chain t-PA.
Quality
Purity: >90% (enzymatic)
Contaminant: <0.5% free 4-nitraniline
Sequence
N-Methylsulfonyl-D-Phe-Gly-Arg-4-nitranilide-acetate
Preparation Note
Working concentration: Approximately 0.25 mM
Storage conditions (working solution): 2 to 8 °C
An aqueous solution of Chromozym t-PA (4mM) is stable for at least two weeks if stored at 2 to 8 °C.Other NotesFor life science research only. Not for use in diagnostic procedures.Legal InformationChromozym is a registered trademark of Pentapharm AG, BaselПараметрыQuality Level 100, assay 90% (enzymatic) form powder mol wt 636.7 packaging pkg of 20 mg Торговая марка Roche concentration 0.25 mM shipped in wet ice storage temp. 20-25°C
Safety InformationRIDADR NONH for all modes of transport WGK Germany WGK 2 Flash Point F Not applicable Flash Point C Not applicable Узнать цену -
Chromozym TH
Кат. номер 11585398001 10206849001 General descriptionChromozym® TH is a chromophoric substrate mostly used to detect thrombin activity†
Tosyl-Gly-Pro-Arg-4-nitranilide acetateApplicationUse Chromozym® TH as a substrate for the determination of serine proteases, specifically thrombin in aqueous solutions.
Quality
Contaminant: 1% free 4-nitraniline
Sequence
Tosyl-glycyl-polyl-arginine-4-nitranilideacetate
Preparation Note
Working concentration: Approximately 0.2 mM
Storage conditions (working solution): 2 to 8 °C
A substrate solution (approx. 2 mM) is stable for at least 4 weeks if stored at 2 to 8 °C.Other NotesFor life science research only. Not for use in diagnostic procedures.Legal InformationChromozym is a registered trademark of Pentapharm AG, BaselПараметрыassay 90% (Tosyl-Gly-Pro-Arg-4-nitranilide acetate (enzymatic)), 90% (enzymatic) Quality Level 100, form powder mol wt Mr 662.62 packaging pkg of 100 mg (11585398001), pkg of 20 mg (10206849001) Торговая марка Roche shipped in ambient storage temp. 20-25°C
Safety InformationRIDADR NONH for all modes of transport Узнать цену -
Chymostatin 11004638001
CAS Number: 9076-44-2 MDL number: MFCD00071059 PubChem Substance ID: 329749270 General descriptionChymostatin is a mixture of three components, A, B, and C. The component A being N-[((S)-1-carboxy-2-phenylethyl)-carbamoyl]--[2-iminohexahydro-4(S)-pyrimidyl]-L-glycyl-L-leucyl-phenylalaninal. The other two components B and C differ in that the L-leucyl residue is substituted by L-valine and L-isoleucine, respectively.ApplicationChymostatin is a specific inhibitor of α-, β-, γ-, and δ-chymotrypsin.
Biochem/physiol Actions
Chymostatin is a strong inhibitor of many proteases, including chymotrypsin, papain, chymotrypsin-like serine proteinases, chymases, and lysosomal cysteine proteinases such as cathepsins A,B,C, B, H, and L. It weakly inhibits human leucocyte elastase. It is effective at a final concentration of 100 to 200 μg/ml (10 to 100 μM). Chymostatin is often included in protease inhibitor cocktails used with plant extracts.
Quality
Performance tested.
Formula variant
C31H41N7O6
Preparation Note
Working concentration: 6 to 60 µg/ml (10 to 100 µM)
1 U chymotrypsin is inhibited to 49% of the original activity by 1.8 µg of chymostatin.
Thin-layer chromatography: butanol/methanol/H2O = 4 / 1 / 2
Working solution: Soluble in glacial acetic acid or DMSO to 20 mg/ml. Sparingly soluble in water, methanol, or ethanol. Insoluble in ethyl acetate, petroleum and ethyl ethers, hexane, or chloroform (CHCl3).
It is recommended to dissolve the inhibitor in 1% acetic acid in higher concentration and to adjust the concentration wanted with phosphate buffer, 0.05 M, pH 7.0, which is common for chymotrypsin assay.
CAUTION: DMSO (Dimethyl sulfoxide) will permeate the skin, carrying solubilized protease inhibitors. Always wear appropriate protection for eyes, skin, etc.
Storage conditions (working solution): -15 to -25 °C
Dilute solutions should be stored frozen in aliquots at -15 to -25 °C and are stable for approximately one month. Avoid repeated freezing. Growth of microorganisms should be avoided as proteases from microbial origin may hydrolyze the peptides.
Solubility testing in glacial acetic acid at 10 mg/ml yields a clear solution, which is usually colorless, but can be yellow in appearance. It is reportedly also soluble in DMSO; only slightly soluble in water and short-chain alcohols; insoluble in ethyl acetate, butyl acetate, ether, hexane, and petroleum ether. Stock solutions (10 mM) can be prepared in DMSO and are stable for months at -20 °C. Stock solutions can also be made in 0.1 M HCl. Dilute solutions (10-100 μM) are only stable for several hours, due to oxidation of the terminal aldehyde.Other NotesFor life science research only. Not for use in diagnostic procedures.Параметрыform powder Quality Level 100, mol wt Mr = 607.71 packaging pkg of 10 mg Торговая марка Roche mp 205 °C solubility acetic acid: soluble 20 mg/mL shipped in wet ice storage temp. 2-8°C SMILES string OC(C(NC(NC(C1NC(NCC1)=N)C([F,Cl,Br,I]C)=O)=O)CC2=CC=CC=C2)=O InChI key MRXDGVXSWIXTQL-HYHFHBMOSA-N
Safety InformationRIDADR NONH for all modes of transport WGK Germany WGK 1 Flash Point F Not applicable Flash Point C Not applicable Узнать цену -
Chymotrypsin Sequencing Grade 11418467001
Enzyme Commission (EC) Number: 3.4.21.1 ( BRENDA | IUBMB ) General descriptionChymotrypsin Sequencing Grade is isolated as a specific protease in pure form from bovine pancreas.
Specificity
Serine endopeptidase that specifically hydrolyzes peptide bonds at the C-terminial of Tyr, Phe, and Trp. Leu, Met, Ala, Asp, and Glu are cleaved at a lower rate. Acts also upon amides and esters of susceptible amino acids. The specificity of Chymotrypsin Sequencing Grade is tested with melittin as a substrate.ApplicationUse Chymotrypsin Sequencing Grade for the hydrolysis of proteins by chymotrypsin alone or in combination with other proteases. It is suitable for peptide mapping, fingerprinting, and sequence analysis.
Quality
Purity: Free of impurities that may interfere with the specific cleavage or separation of peptides in reversed-phase HPLC.
Preparation Note
Working concentration: The recommended amount of enzyme is 1/200 to 1/20 of the quantity of protein by weight.
Storage conditions (working solution): 2 to 8 °C
A solution in 1 mM HCl can be stored for up to one week at 2 to 8 °C.
Inhibitors: Aprotinin, DFP, PMSF, phenothiazine-N-carbonyl chloride, TPCK, ZPCK, 2-macroglobulin, 1-antitrypsin, soybean trypsin inhibitor, and chymostatin. No inhibition by APMSF.
Reconstitution
Dissolve lyophilized chymotrypsin sequencing grade in 1mM HCl. The proteins to be sequenced are dissolved in digestion buffer (100mM Tris-HCl, 10mM CaCl2, pH 7.8).Other NotesFor life science research only. Not for use in diagnostic procedures.ПараметрыQuality Level 100, assay 90% form lyophilized (salt-free) specific activity 70 units/mg protein (at 25 °C, with ATEE as a substrate) mol wt Mr 25 kDa packaging pkg of 4 x 25 µg Торговая марка Roche optimum pH 7.0-9.0 shipped in wet ice storage temp. 2-8°C
Safety Informationpictograms GHS07,GHS08 signalword Danger hcodes H315 - H319 - H334 - H335 pcodes P261 - P264 - P280 - P284 - P304 + P340 + P312 - P342 + P311 Target Organs Respiratory system RIDADR NONH for all modes of transport WGK Germany WGK 1 Flash Point F Not applicable Flash Point C Not applicable Узнать цену -
Citrate Lyase (CL) 10354074001
Enzyme Commission (EC) Number: 4.1.3.6 ( BRENDA | IUBMB ) General descriptionCitrate lyase is considered as a substrate- induced enzyme which helps in catalyzing reversible aldol cleavage of citrate to oxaloacetate and acetate. The enzyme is adaptive to anaerobic dissimilation of citrate.ApplicationIt was used in enzymatic assay to measure citrate concentration using microtiter plates.
Quality
Contaminants: <0.05% ICDH (NAD specific) and “NADH oxidase”, each
Physical form
Lyophilizate, stabilized with BSA, sucrose, MgSO4 and EDTA. Note: A solution in double-distilled water has a pH-value of approximately 7.0. The enzyme can be relyophilized.
Preparation Note
Activator: Mg2+ (Km = 3.0 mM, pH 7.4, 25 °C); other divalent metal ions (Mn2+, Fe2+, Zn2+)Other NotesFor life science research only. Not for use in diagnostic procedures.ПараметрыQuality Level 100, form lyophilized specific activity 0.25 U/mg (At 25 °C with citrate as the substrate.) packaging pkg of 120 U Торговая марка Roche optimum pH 8.0-9.0 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 Узнать цену -
Colcemid 10295892001
Empirical Formula (Hill Notation): C21H25NO5 CAS Number: 477-30-5 Molecular Weight: 371.43 Beilstein/REAXYS Number: 2822892 MDL number: MFCD00075459 PubChem Substance ID: 329749009 General descriptionColcemid is also known as demecolcine. Its generic name is N-methyl-N-deacetyl-colchicine. Colcemid depolymerizes microtubules and blocks mitosis at metaphase.ApplicationColcemid inhibits the formation of mitotic spindles. It is used to increase the percentage of metaphase cells for chromosome analysis.
Biochem/physiol Actions
Often in karyotyping and cell cycle research it is desirable to increase the yield of mitotic cells in a particular phase of the cell cycle. This can be achieved in a variety of ways with the most popular being the use of a cell cycle synchronizing agent such as demecolcine. Demecolcine will arrest cells in metaphase with no remarkable effect on the biochemical events in mitotic cells or in synchronized G1 and S phase cells. White blood cells are often treated with demecolcine to arrest cells in metaphase.
Physical form
Solution (10 μg/ml), filtered through 0.2 μm pore size membrane.Other NotesFor life science research only. Not for use in diagnostic procedures.ПараметрыQuality Level 100, sterility sterile; 0.2 µm filtered form solution packaging pkg of 20 mL (10 µg/ml) Торговая марка Roche application(s) blocking: suitable impurities microbial, tested solubility water: miscible shipped in wet ice storage temp. 2-8°C SMILES string CN[C@H]1CCc2cc(OC)c(OC)c(OC)c2C3=CC=C(OC)C(=O)C=C13 InChI 1S/C21H25NO5/c1-22-15-8-6-12-10-18(25-3)20(26-4)21(27-5)19(12)13-7-9-17(24-2)16(23)11-14(13)15/h7,9-11,15,22H,6,8H2,1-5H3/t15-/m0/s1 InChI key NNJPGOLRFBJNIW-HNNXBMFYSA-N
Safety InformationRIDADR NONH for all modes of transport WGK Germany nwg Flash Point F does not flash Flash Point C does not flash Узнать цену -
Collagen 11179179001
General descriptionCollagens are abundantly present in vertebrates. 41 specific genes codes for 42 polypeptide chains and produce 27 collagen types. Collagens are mainly present in the extracellular matrix. Collagen type I is abundantly present in mammals. Type IV collagen is an important structural protein of basement membranes. It is chemically and genetically different from stroma collagen types I and III and cartilage collagen type II.
Specificity
Active on most mammalian cells.ApplicationCollagen is used as a substrate for culturing a variety of cells, such as coating of culture dishes and the preparation of collagen gels.
Collagen has been used in the preparation of coverslip coating solution and collagen gel contraction assay.
Biochem/physiol Actions
Collagen provides extracellular support for multicellular animals. Collagen type I offers mechanical stability, strength and toughness to a range of tissues from tendons and ligaments, to skin, cornea, bone and dentin.
Features and Benefits
Contents
Lyophilizate, cell culture grade, 3 vials of 10mg
Specifications
Biological activity: Tested for the promotion of adherence of HUV-EC cells.
Preparation Note
Working concentration: 5 µg/cm2
5 µg/cm2 for the coating of cell culture vessels; for the preparation of collagen gels, a final concentration of 2 to 3 mg/ml is used.
Reconstitution
IFor best results, dissolve the lyophilizate in 0.2% acetic acid (v/v) solution.
For the preparation of collagen gels, the content of the bottle should be dissolved in 3.3 ml sterile 0.2% acetic acid (v/v) each. This results in a final concentration of 3 mg/ml.
For coating culture dishes the final concentration should be 1 to 2 mg/ml.
Note: For dissolving: do not stir, just pour the acetic acid onto the lyophilizate and allow it to stand for several hours until it has dissolved. To completely dissolve the product an incubation for up to a maximum of 24 hours at 15 to 25 °C may be required.
Reconstitution of the Collagen should be done at 15 to 25 °C, higher temperatures destroy the fibres and may prevent the subsequent gelation of the Collagen.Other NotesFor life science research only. Not for use in diagnostic procedures.Параметрыbiological source rat tail Quality Level 100 sterility sterile form lyophilized (clear, colorless solution after reconstitution) packaging pkg of 30 mg Торговая марка Roche application(s) cell culture | mammalian: suitable UniProt accession no. P02454, shipped in wet ice storage temp. 2-8°C Gene Information rat ... Col1a1(29393)
Safety InformationRIDADR NONH for all modes of transport WGK Germany WGK 1 Flash Point F does not flash Flash Point C does not flash Узнать цену -
Collagenase B
Enzyme Commission (EC) Number: 3.4.24.3 ( BRENDA | IUBMB )
Кат. номер 11088807001 11088831001 11088815001 General descriptionCollagenase B is prepared from Clostridium histolyticum cultures by filtration, ammonium sulfate precipitation, dialysis, and lyophilization. Collagenase from Clostridium histolyticum is a collagenolytic enzyme. It interacts with collagen-like substances and is responsible for the degradation of collagen into small peptides. It is widely used for the disaggregation of many types of tissues (e.g., lung, heart, muscle, bone, adipose tissue, liver, kidney, cartilage, mammary gland, placentae, blood vessels, brain, tumors) and for the preparation of single-cell suspensions for the establishment of primary cell culture systems. Collagenase B is recommended when yield and viability are important.
Specificity
Collagenase degrades native collagen. Clostripain, trypsin-like enzymes, and neutral proteases degrade other proteins as well.ApplicationClostridium collagenase from Roche has been used to prepare cells from many types of tissue, such as hepatocytes, adipocytes, pancreatic islets, epithelial cells, muscle cells, endothelial cells, etc. However, suitability of each lot of the enzyme for disruption of a particular tissue should be determined empirically.
Collagenase B has been used for preparation of cells from mouse aortas and colorectal cancer tissue.
Specifications
Enzyme activity:
Collagenase activity: >0.15U/mg (according to Wunsch) (+25°C, 4-phenyl-azobenzyl-oxycarbonyl-Pro-Leu-Gly-Pro-D-Arg as the substrate)
Contaminating enzyme activities: trypsin, clostripain, and total proteolytic activity
Collagenase B has a normal to high collagenase activity and a higher than average clostripain activity (usually >10U/mg).
Inhibitors:
Collagenase inhibitors: EDTA, EGTA, Cys, His, DTT, 2-mercaptoethanol
Collagenase is not inhibited by serum.
Clostripain inhibitors: TLCK
Trypsin inhibitors: aprotinin, trypsin inhibitor (egg white, soybean), serum
Unit Definition
Collagenase from Roche is assayed in Wnsch units (1 µmol of product formed per minute at +25 °C with Wnsch substrate).
Frequently, collagenase activities are given in Mandl units (1 µmol leucine liberated from collagen in 5 hours at +37 °C).
Unfortunately, there is no consistent conversion factor between the two units of activity, since the Mandl unit depends, in part, on the concentration of contaminating proteases in the collagenase preparation, an indefinable variable. A purer collagenase preparation would actually give a lower specific activity in Mandl units than a crude preparation. Clostridium preparations typically give conversion factors of approximately 1:1800 (e.g., a particular lot of Clostridium collagenase contained approximately 0.15 Wnsch U/mg and 250 Mandl U/mg).
Preparation Note
Activator: Ca2+
Working concentration: 0.5 to 2.5 mg/ml
Storage conditions (working solution): -15 to -25 °C
Roche recommends reconstituting only the amount of lyophilizate needed for immediate use. The reconstituted solution can be stored at -15 to -25 °C for up to one week. Avoid repeated freezing and thawing since activity decreases after reconstitution.
Reconstitution
Reconstitution in any balanced salt solution (e.g., HBSS)Other NotesFor life science research only. Not for use in diagnostic procedures.ПараметрыQuality Level 100, sterility non-sterile form lyophilized packaging pkg of 100 mg (11088807001), pkg of 2.5 g (11088831001), pkg of 500 mg (11088815001) Торговая марка Roche concentration 0.5-2.5 mg/mL optimum pH 6.0-8.0 shipped in wet ice storage temp. 2-8°C
Safety InformationRIDADR NONH for all modes of transport Узнать цену -
Collagenase D
Enzyme Commission (EC) Number: 3.4.24.3 ( BRENDA | IUBMB )
Кат. номер 11088858001 11088882001 11088866001 General descriptionCollagenase D is prepared from Clostridium histolyticum cultures by filtration, ammonium sulfate precipitation, dialysis, and lyophilization.SpecificityCollagenase degrades native collagen. Clostripain, trypsin-like enzymes, and neutral proteases degrade other proteins as well.ApplicationCollagenase from C. histolyticum is widely used for the disaggregation of many types of tissues (e.g., lung, heart, muscle, bone, adipose tissue, liver, kidney, cartilage, mammary gland, placentae, blood vessels, brain, tumors) and for the preparation of single cell suspensions for the establishment of primary cell culture systems. Collagenase D is recommended when functionality and integrity of cell-surface proteins are important.
Clostridium collagenase from Roche has been used to prepare cells from many types of tissue, such as hepatocytes, adipocytes, pancreatic islets, epithelial cells, muscle cells, endothelial cells, etc. However, suitability of each lot of the enzyme for disruption of a particular tissue should be determined empirically.Features and BenefitsContents
Lyophilizate, nonsterileUnit DefinitionCollagenase from Roche is assayed in Wnsch units (1 µmol of product formed per minute at +25 °C with Wnsch substrate).
Frequently, collagenase activities are given in Mandl units (1 µmol leucine liberated from collagen in 5 hours at +37 °C).
Unfortunately, there is no consistent conversion factor between the two units of activity, since the Mandl unit depends, in part, on the concentration of contaminating proteases in the collagenase preparation, an indefinable variable. A purer collagenase preparation would actually give a lower specific activity in Mandl units than a crude preparation. Clostridium preparations typically give conversion factors of approximately 1:1800 (e.g., a particular lot of Clostridium collagenase contained approximately 0.15 Wnsch U/mg and 250 Mandl U/mg).Physical formLyophilizate, nonsterile.Preparation NoteActivator: Ca2+
Working concentration: 0.5 - 2.5mg/ml
Storage conditions (working solution): -15 to -25°C
Roche recommends reconstituting only the amount of lyophilizate needed for immediate use. The reconstituted solution can be stored at -15 to -25°C for up to one week. Avoid repeated freezing and thawing since activity decreases after reconstitution.
Inhibitors:
Collagenase inhibitors: EDTA, EGTA, Cys, His, DTT, 2-mercaptoethanol
Collagenase is not inhibited by serum.
Clostripain inhibitors: TLCK
Trypsin inhibitors: aprotinin, trypsin inhibitor (egg white, soybean)
Enzyme activity:
Collagenase activity: >0.15 U/mg (according to Wunsch) (+25°C, 4-phenyl-azobenzyl-oxycarbonyl-Pro-Leu-Gly-Pro-D-Arg as the substrate)
Contaminating enzyme activities: trypsin, clostripain, and total proteolytic activity
Collagenase D has a normal to high collagenase activity and very low tryptic activity (usually <0.2 units/mg, BAEE as substrate).ReconstitutionReconstitution in any balanced salt solution (e.g., HBSS)Other NotesFor life science research only. Not for use in diagnostic procedures.ПараметрыQuality Level 100 sterility non-sterile form lyophilized packaging pkg of 100 mg (11088858001), pkg of 2.5 g (11088882001), pkg of 500 mg (11088866001) Торговая марка Roche concentration (working concentration: 0.5 - 2.5 mg/ml) optimum pH 6.0-8.0 shipped in wet ice storage temp. 2-8°C
Safety InformationRIDADR NONH for all modes of transport Узнать цену -
Collagenase H
Enzyme Commission (EC) Number: 3.4.24.3 ( BRENDA | IUBMB )
Кат. номер 11074032001 11087789001 11074059001 General descriptionCollagenase H is an enzyme mixture prepared from Clostridium histolyticum cultures by filtration, ammonium sulfate precipitation, dialysis, and lyophilization.
Specificity
Collagenase degrades native collagen. Clostripain, trypsin-like enzymes, and neutral proteases also degrade other proteins.ApplicationCollagenase from C. histolyticum is used for the dissociation of tissues for the establishment of primary cell cultures. Collagenase H is well suited for the isolation of hepatocytes from rat liver by the collagenase perfusion method. This enzyme preparation is also suitable for the preparation of other types of cells such as endothelial cells from large vessels, and for the isolation of adipocytes from epididymal fat pads of rats.
Physical form
Lyophilizate, nonsterile
Preparation Note
Enzyme activity:
Collagenase activity: >0.15U/mg (according to Wunsch) (+25°C, 4-phenyl-azobenzyl-oxycarbonyl-Pro-Leu-Gly-Pro-D-Arg as the substrate).
Contaminating enzyme activities: trypsin, clostripain, and total proteolytic activity
Collagenase H has a balanced ratio of enzyme activities, and is function tested for the isolation of rat hepatocytes (perfusion method).
Inhibitors:
Collagenase inhibitors: EDTA, EGTA, Cys, His, DTT, 2-mercaptoethanol
Collagenase is not inhibited by serum.
Clostripain inhibitors: TLCK
Trypsin inhibitors: aprotinin, trypsin inhibitor (egg white, soybean)
Activator: Ca2+
Working concentration: 0.5 to 2.5mg/ml,
Approximately 1 mg/ml for the isolation of rat hepatocytes and 2mg/ml for the isolation of adipocytes.
Storage conditions (working solution): -15 to -25°C
The reconstituted solution should be stored at -15 to -25°C for short term storage, -60°C or as described below for long-term storage.
Reconstitution
Reconstitution in any balanced salt solution (e.g., HBSS)Other NotesFor life science research only. Not for use in diagnostic procedures.ПараметрыQuality Level 100, sterility non-sterile form lyophilized packaging pkg of 100 mg (11074032001), pkg of 2.5 g (11087789001), pkg of 500 mg (11074059001) Торговая марка Roche optimum pH 6.0-8.0 shipped in wet ice storage temp. 2-8°C
Safety InformationRIDADR NONH for all modes of transport Узнать цену -
cOmplete™ His-Tag Purification Column
Кат. номер 6781543001 6781535001 General descriptionThe most common technique in affinity purification of proteins involves engineering a sequence of 6 to 14 histidines into the N- or C-terminal (or even on an exposed loop) of the protein. Such polyhistidine stretches bind strongly to divalent metal ions such as nickel and cobalt. This effect can be used to separate proteins. Metal ions can be immobilized on a matrix using a chelator, which still allows the ion to interfere with the polyhistidine tag of the protein. When these his-tagged proteins are passed through a column containing immobilized metal ions, the proteins bind via the tag to the column. Nearly all untagged proteins pass through the column. The protein is released from the column by elution with either imidazole, which competes with the polyhistidine tag for binding to the column, or by a decrease in pH, which decreases the affinity of the tag for the resin. While this procedure is generally used for the purification of recombinant proteins with an engineered affinity tag, it can also be used for natural proteins with an inherent affinity for divalent cations.
Features and Benefits
cOmplete His-Tag Purification Columns are available in 2 sizes and prepacked with cOmplete His-Tag Purification Resin. The cOmplete His-Tag Purification Resin is an innovative high-capacity IMAC matrix (Immobilized Metal Affinity Chromatography) for convenient single-step purifications of His-tagged proteins from total lysates. Roches propriety nickel-chelate chemistry ensures extraordinary compatibility with commonly used reducing agents such as DTT, chelating metalloprotease inhibitors such as EDTA, and a wide range of buffer substances and salt conditions. The wide choice of compatible ingredients allows optimization of buffers for maximum protein stability and solubility.
cOmplete His-Tag Purification Columns are fully compatible with standard purification systems such as KTA Systems (Cytiva).
- Use the buffer conditions best suited to your protein
- Repeatedly obtain highly pure protein
- Protect your protein from toxic nickel
- Work in a safe and eco-friendly environment
Packaging- 06781535001 (5 mL column; 1 column containing 5 ml of cOmplete His-Tag Purification Resin)
- 06781543001 (5 x 1 mL columns; 5 columns containing 1 ml of cOmplete His-Tag Purification Resin)
Compatibility
Compatibility for long term storage: 20% ethanol, pH 4.0 to pH 9.0
Compatibility during chromatography: The resin is compatible with 10mM EDTA, 10mM DTT during the purification (1 hour incubation), 6M guanidinium-HCl, 8M urea, pH 2.0 to pH 14.0.
Compatibility during cleaning: 4% SDS Form cOmplete His-Tag Purification Resin filled in columns, pre-charged with Ni2+ stored in 20% ethanol.Other NotesRecommended volumetric flow rate: 5 ml column (06 781 535 001): 2.5 to 10 ml/minute
1 ml column (06 781 543 001): 0.5 to 2.0 ml/minute
The volumetric flow rate is a function of the columns cross section.
Recommended imidazole concentration for load/wash: Nonspecific binding of proteins without a His-tag is low.
Use up to 5 mM imidazole in load and/or wash buffers.
If establishing a new assay for purification of His-tag proteins with cOmplete His-Tag Purification Columns do not use imidazole. If, e.g., the purity of the His-tag protein needs to be improved following this first step, use imidazole in a final concentration of up to 5mM in a second step.
Recommended imidazole concentration for elution: Up to 500mM
Please note:
In contrast to other available resins, bound His-tagged protein typically elutes from cOmplete His-Tag Purification Columns with a lower imidazole concentration, e.g., 25 to 45mM.
Binding capacity
The binding capacity of the resin to various types of proteins may vary according to the protein characteristics such as the size of the protein.
cOmplete His-Tag Purification Columns bind with a high specificity to the polyhistidine-tagged protein. As a consequence, the binding kinetics may appear to be different when compared to conventional metal chelate matrices. Full capacity of cOmplete His-Tag Purification Columns can be achieved by allowing more time for the protein to bind to the resin by lowering the flow rate during the chromatography purification procedure.
For life science research only. Not for use in diagnostic procedures.Legal InformationcOmplete is a trademark of RocheПараметрыpackaging pkg of 5 x 1 mL (columns (06781543001)), pkg of 5 mL (column (06781535001)) Торговая марка Roche parameter 1,420 cm/hr max. flow rate capacity 40 mg/mL, resin binding capacity shipped in wet ice storage temp. 2-8°C
Safety InformationRIDADR NONH for all modes of transport Узнать цену -
cOmplete™ His-Tag Purification Resin
Кат. номер 5893682001 5893801001 General descriptionBead size: 45 to 165µM
Binding capacity: 40mg protein per ml bed volume of resin. The binding capacity of the resin to various types of proteins may vary according to the protein characteristics such as the size of the protein. cOmplete His-Tag Purification Columns bind with a high specificity to the polyhistidine-tagged protein. As a consequence, the binding kinetics may appear to be different when compared to conventional metal chelate matrices. Full capacity of cOmplete His-Tag Purification Columns can be achieved by allowing more time for the protein to bind to the resin by lowering the flow rate during the chromatography purification procedure.
Maximal linear flow rate: 1,420cm/hour
Recommended volumetric flow rate: The volumetric flow rate is a function of the cross section of the column. Using the following formula, a linear flow rate can be converted to a volumetric flow (mL/min.): Linear flow rate (cm/hour) x column cross sectional area (cm2)/60. The column cross sectional area is defined as x r2, whereas is the constant pi and r is the inner radius of the column.
Recommended imidazole concentration for load/wash: Nonspecific binding of proteins without a His-tag is low. Use up to 5mM imidazole in load and/or wash buffers. If establishing a new assay for purification of His-tag proteins with cOmplete His-Tag Purification Columns, do not use imidazole. If, e.g., the purity of the His-tag protein needs to be improved following this first step, use imidazole in a final concentration of up to 5mM in a second step.
Recommended imidazole concentration for elution: Up to 500 mM. Please note: Contrary to other available resins, bound His-tagged protein typically elutes from cOmplete His-Tag Purification Columns with a lower imidazole concentration, e.g., 25 to 45mM.
Compatibility for long term storage: 20% ethanol, pH 4.0 to pH 9.0
Compatibility during chromatography: The resin is compatible with 10mM EDTA, 10mM DTT during the purification (1 hour incubation), 6M guanidinium-HCl, 8M urea, pH 2.0 to pH 14.0.
Compatibility during cleaning: 4% SDS
The cOmplete His-Tag Purification Resin is an innovative high-capacity IMAC matrix (Immobilized Metal Affinity Chromatography) for the purification of histidine-tagged proteins via batch or liquid chromatography procedures from total lysates. In contrast to all currently available resins, which are primarily based on NTA or IDA chelator chemistry, this resin is the only resin on the market which tolerates buffers that contain EDTA. Because EDTA is a known inhibitor of metalloproteases–which are frequently present in any cell type–this chromatography material provides the option to increase the protection of your target protein by supplementing your buffers with EDTA. This resin therefore allows increased protection of your proteins against degradation and, at the same time, enriches the protein using the well-established affinity purification method.
In addition, the resin maintains its binding capacity whether you are using low- or high-molecular weight proteins, and its specificity allows a one-step purification. By using one of the strongest chelators available, minimal contamination of your flow-through fractions with metal ions is ensured, safeguarding your downstream applications.
The cOmplete His-Tag Purification Resin is also available as prepacked format: with 1mL or 5mL resin.ApplicationRecombinant Protein Expression
Purifying a protein of interest is often essential for determining its function, structure, or interactions, for raising specific antibodies, or preparing enzymes for practical applications. Isolation of naturally expressed proteins from their original source can be a complex process involving numerous chromatographic steps. Recombinant protein expression in dedicated host organisms can greatly simplify this task. Such expression systems generally ensure higher expression levels. Fusing the target protein to a tag also confers advantageous binding ability to an affinity matrix.
Protein Purification using Immobilized Ni2+
The most common technique in affinity purification of proteins involves engineering a sequence of 6 to 14 histidines into the N- or C-terminal (or even on an exposed loop) of the protein. Such polyhistidine stretches bind strongly to divalent metal ions such as nickel and cobalt. This effect can be used to separate proteins. Metal ions can be immobilized on a matrix using a chelator, which still allows the ion to interfere with the polyhistidine tag of the protein. When these his-tagged proteins are passed through a column containing immobilized metal ions, the proteins bind via the tag to the column. Nearly all untagged proteins pass through the column. The protein is released from the column by elution with either imidazole, which competes with the polyhistidine tag for binding to the column, or by a decrease in pH, which decreases the affinity of the tag for the resin. While this procedure is generally used for the purification of recombinant proteins with an engineered affinity tag, it can also be used for natural proteins with an inherent affinity for divalent cations.
His-Tags
Ideally, the His-tagged target protein binds much stronger to the Ni2+ chelate matrix than endogenous histidine-containing protein of the expression host. Relative binding strength depends on how many histidines can bind simultaneously to the matrix (avidity effect). Longer His-tags confer stronger binding and better separation of the target from potentially contaminating host proteins. The classic His-tag has six consecutive histidines. Tags with 10 to 14 histidines may produce a better purification. Most importantly, His-tagged proteins can be purified using Ni2+ chelate matrices under both native and denaturing conditions. Due to their hydrophilic and flexible nature, these matrices increase the solubility of the target proteins and only rarely interfere with protein function. This unique combination of features enables the His-tag to be a versatile tool for a wide range of protein purification applications.
Features and Benefits
cOmplete His-Tag Purification Resin is the only resin for purifying large amounts of protein without compromises.
- Use the buffer conditions best suited to your protein: Keep your protein comfortable and let it, not your purification resin, determine whether you use DTT, EDTA, or other buffer substances.
- Repeatedly obtain highly pure protein: Single step purification without resin recharging.
- Protect your protein from toxic nickel: Reduce protein oxidation and aggregation caused by resins that leach nickel.
- Work in a safe and eco-friendly environment: Avoid handling of toxic nickel and completely eliminate disposal costs.
Physical form
50% resin in suspension, pre-charged with Ni2+Other NotesFor life science research only. Not for use in diagnostic procedures.
EDTA-compatible resin for the purification of poly-histidine-tagged proteins.Legal InformationcOmplete is a trademark of RocheПараметрыpackaging pkg of 200 mL (05893801001 settled resin volume), pkg of 25 mL (05893682001 settled resin volume) mfr. no. Roche matrix Sepharose-CL 6B shipped in ambient storage temp. 2-8°C
Safety Information
