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G-418 Solution
Empirical Formula (Hill Notation): C20H40N4O10 · 2H2SO4 CAS Number: 108321-42-2 Molecular Weight: 692.71 MDL number: MFCD00058314 PubChem Substance ID: 329799925
Кат. номер 4727894001 4727878001 General descriptionO-2-Amino-2,7-dideoxy-δ-glycero-α-δ-glucoheptopyranosyl[1->4]-O-3-deoxy-4C-methyl-3-[methyl-amino]-β-L-arabinopyranosyl-δ-streptamine, disulfate salt G-418, an aminoglycoside antibiotic, is used as a dominant selection agent in transfection experiments. The antibiotic interferes with the function of 80S ribosomes and blocks protein synthesis in eukaryotic cells. Transfection of neomycin-resistance genes into cells results in resistance to G-418 and enables the cells to grow in culture media containing G-418.ApplicationG-418 Solution is used to select eukaryotic cells that are stably transfected with the neomycin resistance gene (neo), and to maintain the phenotype (neor) of resistant cells.
G-418 is also used to eliminate contaminating fibroblasts from mixed cultures.
Biochem/physiol Actions
Mode of Action: G418 blocks polypeptide synthesis and protein elongation by inhibiting synthesis at the 70S and 80S ribosomes.
Antimicrobial Spectrum: G418 selects for cells stably transfected with an iNOS promoter construct and neomycin resistance gene and shows activity against protozoa and helminthes.
Features and Benefits- Convenient sterile-filtered solution for immediate use – saves valuable time and effort, and avoids potential errors arising from the preparation of sterile working stock solution from G-418 powder.
- Consistent quality and potency – shows no lot-to-lot variation in its ability to select resistant cells.
- Function tested in a cell culture assay - ensures effectiveness.
- Very pure, as indicated by TLC and microbiological assays – allows you to use less antibiotic while producing greater selection pressure. Lack of contaminants means minimal toxicity.
- Cost effective, since the selected colonies are stable.
Aqueous solution of G-418, 892 µg/mg, filtered through 0.2µm pore size membrane
Quality
Purity: = 98%, as shown by TLC.
Function testing: in cell culture.
Preparation Note
Working concentration: The optimal concentration of G-418 must be determined experimentally and varies with the cell type used.
Active weight: >700µg/mg
Recommended concentration in cell culture: The optimal concentration of G-418 must be determined experimentally; it varies from 100µg/ml to 1mg/ml, depending on the cell type used.
Specific rotation: 104° to 121°, in water (on dry basis, according to USP XXII).Other NotesFor life science research only. Not for use in diagnostic procedures.Параметрыassay 98% (TLC) Quality Level 100, form solution mol wt 692.7 Da packaging pkg of 100 ml (04727894001 [5 x 20 ml, equivalent to 5 g]), pkg of 20 mL (04727878001 [equivalent to 1 g]) Торговая марка Roche concentration 50 mg/mL application(s) transfection: suitable Mode of action protein synthesis | inhibits antibiotic activity spectrum protozoa shipped in wet ice storage temp. 2-8°C SMILES string OS(O)(=O)=O.OS(O)(=O)=O.CN[C@H]1[C@H](O)[C@@H](OC[C@@]1(C)O)O[C@H]2[C@H](N)C[C@H](N)[C@@H](O[C@H]3O[C@H]([C@H](C)O)[C@@H](O)[C@H](O)[C@H]3N)[C@@H]2O InChI 1S/C20H40N4O10.2H2O4S/c1-6(25)14-11(27)10(26)9(23)18(32-14)33-15-7(21)4-8(22)16(12(15)28)34-19-13(29)17(24-3)20(2,30)5-31-19;2*1-5(2,3)4/h6-19,24-30H,4-5,21-23H2,1-3H3;2*(H2,1,2,3,4)/t6?,7-,8+,9+,10+,11-,12-,13-,14+,15+,16-,17-,18+,19-,20+;;/m0../s1 InChI key UHEPSJJJMTWUCP-NKCAIAFTSA-N
Safety InformationRIDADR NONH for all modes of transport -
4743784001
GC-RICH PCR System, dNTPack 4743784001
General descriptionThe GC-RICH PCR System, dNTPack is composed of a special enzyme blend of thermostable Taq DNA Polymerase and Tgo DNA Polymerase, a thermostable enzyme with a proofreading (3′-5′ exonuclease) activity. This polymerase mixture by itself outperforms Taq DNA Polymerase in respect to yields, fidelity and specificity beside the possibility to amplify fragments up to 5 kb in length. The GC-RICH PCR reaction buffer in combination with the included GC-RICH resolution solution allows to amplify very efficiently difficult templates like GC-rich targets.ApplicationThe GC-RICH PCR System, a blend of Taq DNA Polymerase and a proofreading polymerase, enables amplification of templates that are difficult or impossible to amplify with other polymerases and other blends of polymerases. The enhanced processivity of the blend and the unique GC-RICH Resolution Solution combine to deliver superior performance – especially from problematic templates.
The GC-RICH PCR System, dNTPack may also be used in standard PCR applications, providing improved results (higher yield, higher accuracy) over Taq DNA Polymerase alone.
Features and Benefits- Ease access to difficult templates:-
- Reagents, the GC-RICH Resolution Solution and PCR Grade Water are provided.
- Amplify DNA fragments up to 5 kb.
- Cost-effective:-
Packaging
1 kit containing 6 components
Quality
The GC-RICH PCR System is function-tested, by amplifying a human 284 bp ApoE genomic fragment, using the GC-RICH PCR System protocols.
Unit Definition
Volume Activity: 2 U/μl
Preparation Note
Working 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.Other NotesFor life science research only. Not for use in diagnostic procedures.Legal InformationUse of this product is covered by US patent claims and corresponding patent claims outside the US. The purchase of this product includes a limited, non-transferable immunity from suit under the foregoing patent claims for using only this amount of product for the purchaser′s own internal research. No right under any other patent claim, no right to perform any patented method, and no right to perform commercial services of any kind, including without limitation reporting the results of purchaser′s activities for a fee or other commercial consideration, is conveyed expressly, by implication, or by estoppel. This product is for research use only. Diagnostic uses under Roche patent claims require a separate license from Roche. Further information on purchasing licenses may be obtained by contacting the Director of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA.Параметрыusage sufficient for ≤50 reactions feature Difficult Templates/Specialty Enzymes PCR, dNTPs included, hotstart: no packaging pkg of 100 U Торговая марка Roche parameter 72 °C optimum reaction temp. application(s) PCR: suitable input purified DNA shipped in dry ice storage temp. −20°C (−15°C to −25°C)
Safety Informationhcodes H412 pcodes P273 - P501 RIDADR NONH for all modes of transport WGK Germany WGK 2 Flash Point F does not flash Flash Point C does not flash -
4634772001
Genopure Buffer Set 4634772001
General descriptionDoubling the volume of the Suspension, Lysis, and Neutralization Buffers during the Genopure plasmid preparation procedure will provide better yields of plasmid DNA from low-copy number plasmids. The buffers in this set may be used with either of the Genopure Plasmid Kits (Midi or Maxi).ApplicationDoubling the volume of the Suspension, Lysis, and Neutralization Buffers during the Genopure plasmid preparation procedure will provide better yields of plasmid DNA from low-copy number plasmids. The buffers in this set may be used with either of the Genopure Plasmid Kits (Midi or Maxi).
Features and Benefits
The Genopure Buffer Set for Low-Copy Number Plasmids provides additional buffers to supplement those in the Genopure Plasmid Kits.
RNase A is also included in the set because it eliminates bacterial RNA from the preparation, thereby improving the recovery of low copy number plasmids.- Convenient, ready-to-use, function-tested, nuclease-free buffers
- Helps the Genopure Plasmid Midi and Maxi Kits enhance plasmid yield
- Ensures reproducible results
Components- Suspension Buffer, 500 mL
- RNase A, 50 mg
- Lysis Buffer, 500 mL
- Neutralization Buffer, 500 mL
Other NotesFor life science research only. Not for use in diagnostic procedures.
To learn how these buffers are used, see the product listings or the package inserts for the Genopure Plasmid Midi Kit and the Genopure Plasmid Maxi Kit.ПараметрыQuality Level 100, Торговая марка Roche packaging kit of for 20 maxi preps or 60 midi preps
for low copy number plasmids
Safety Informationpictograms GHS05 signalword Danger hcodes H290 - H314 pcodes P280 - P301 + P330 + P331 - P303 + P361 + P353 - P304 + P340 + P310 - P305 + P351 + P338 + P310 - P390 RIDADR UN 1824 8 / PGIII WGK Germany WGK 1 Flash Point F does not flash Flash Point C does not flash -
3143422001
Genopure Plasmid Maxi Kit 3143422001
General descriptionFor large-scale (maxi) preparation of plasmid DNA.ApplicationThe Genopure Plasmid Maxi Kit prepares transfection-grade plasmid DNA in large quantities (up to 500 µg plasmid) from bacterial cultures. Isolated plasmid is suitable for most molecular biology applications:- Transfection
- Southern blotting
- Sequencing
- PCR/long PCR
- Restriction digestion
- Cloning
Features and Benefits
The Genopure Plasmid Maxi Kit prepares highly purified plasmid DNA in large quantities using a modified alkaline lysis method.- Save time with ready-to-use reagents.
- Purify all sizes and types of plasmid
- Process multiple samples in parallel
- Eliminate the use of hazardous organic compounds
- Obtain higher purity plasmid DNA
Components- Suspension Buffer
- RNase A
- Lysis Buffer
- Neutralization Buffer
- Equilibration Buffer
- Wash Buffer
- Elution Buffer
- NucleoBond AX 500 Columns
- Folded Filters (240 mm diameter)
- Sealing Rings
Quality
Plasmid DNA purified by this kit has been tested for restriction digestion; pUC 19 was isolated from transformed HB101 as described in the protocol. 1 µg of plasmid was completely digested with 1 U Msp I for 2 hours at +37°C, as shown by agarose gel analysis.
Plasmid recovery was tested with 250 µg purified plasmid. The recovery was >90%, with more than 80% in supercoiled form. The yield of plasmid DNA was determined by isolating pBS from DH5a cells. From 150 ml culture volume with a density of A600 between 3 and 6, >400 µg of plasmid DNA was obtained.
The purity (checked by the ratio of A260/A280) is 1.8 + 0.2.
RNA contamination was analyzed with 3 µg pBS purified with the standard procedure and checked by electrophoresis on an agarose gel. No RNA was detected.
The kit components have been tested for the absence of nucleases according to current quality control procedures.
Preparation Note
The isolation procedure is based on a modified alkaline lysis protocol and can be divided into the following steps:The bacteria are partially lysed, allowing the plasmid DNA to escape the cell wall into the supernatant. The larger E. coli chromosomal DNA is trapped in the cell wall. The lysate is cleared of cellular debris by filtration or centrifugation, and the plasmid DNA-containing fraction is loaded onto a pre-equilibrated column. Since the cleared lysate is applied to the column in a low-salt buffer, plasmid DNA binds to the macroporous anion-exchange material in the column. Cellular impurities are eluted from the column with a high-salt wash. Finally, the plasmid DNA is eluted from the column. Recovered DNA is precipitated from the eluate to remove salt and concentrate the plasmid.
Analysis Note
Sample:
E. coli culture that contains a high-copy number plasmid: 30 to 150 mL bacterial culture
E. coli culture that contains a low-copy number plasmid: 100 to 500 mL bacterial culture
Plasmid Size: The isolation procedure is suitable for all sizes of plasmid. Note: Lysates of larger constructs (up to 100 kb) should be cleared by filtration rather than centrifugation to avoid shearing the plasmid.
Time Required: 75 minutes (including filtration of the lysate)
Typical Yield:
High-copy number plasmid: 3 to 5 µg/mL culture
Low-copy number plasmid: 0.2 to 1 µg/mL culture
Product Purity: Isolated plasmid DNA is free of other bacterial components, including RNA.Other NotesIon Exchange Chromatography,
For life science research only. Not for use in diagnostic procedures.ПараметрыQuality Level 100, Торговая марка Roche packaging kit of for 10 isolations from 30 to 150 ml
Safety Informationpictograms GHS02,GHS05 signalword Danger hcodes H226 - H290 - H314 pcodes P210 - P280 - P303 + P361 + P353 - P304 + P340 + P310 - P305 + P351 + P338 + P310 - P370 + P378 RIDADR UN 3316 9 WGK Germany WGK 1 Flash Point F 100.4 °F Flash Point C 38 °C -
3143414001
Genopure Plasmid Midi Kit 3143414001
General descriptionPlasmid recovery was tested with 50 µg purified plasmid. The recovery was >90%, with more than 80% in supercoiled form. The yield of plasmid DNA was determined by isolating pBS from DH5a cells. From 30 mL culture volume with a density of A600 between 3 and 6, >85 µg of plasmid DNA was obtained.
The purity (checked by the ratio of A260/A280) is 1.8 + 0.2.
No RNA is detectable.
The kit components have been tested for the absence of nucleases according to current quality control procedures.ApplicationThe Genopure Plasmid Midi Kit prepares transfection-grade plasmid DNA in medium quantities (up to 100 µg plasmid) from bacterial cultures. Isolated plasmid is suitable for most molecular biology applications:- Transfection
- Southern blotting
- Sequencing
- PCR
- Restriction analysis
- Cloning
Features and Benefits
The Genopure Plasmid Midi Kit prepares highly purified plasmid DNA in medium quantities using a modified alkaline lysis method.- Save time with ready-to-use reagents.
- Purify all sizes and types of plasmid,
- Process multiple samples in parallel
- Eliminate the use of hazardous organic compounds
- Obtain higher purity plasmid DNA
Components- Suspension Buffer
- RNase A
- Lysis Buffer
- Neutralization Buffer
- Equilibration Buffer
- Wash Buffer
- Elution Buffer,
- NucleoBond AX 100 Columns
- Folded Filters (150 mm diameter)
- Sealing Rings
Quality
Plasmid DNA purified by this kit has been tested for restriction digestion; pUC 19 was isolated from transformed HB101 as described in the protocol. 1 µg of plasmid was completely digested with 1 U Msp I for 2 hours at +37°C, as shown by agarose gel analysis.
Plasmid recovery was tested with 50 µg purified plasmid. The recovery was >90%, with more than 80% in supercoiled form. The yield of plasmid DNA was determined by isolating pBS from DH5a cells. From 30 ml culture volume with a density of A600 between 3 and 6, >85 µg of plasmid DNA was obtained.
The purity (checked by the ratio of A260/A280) is 1.8 + 0.2.
RNA contamination was analyzed with 3 µg pBS purified with the standard procedure and checked by electrophoresis on an agarose gel. No RNA was detected.
The kit components have been tested for the absence of nucleases according to current Quality Control procedures.
Preparation Note
The isolation procedure is based on a modified alkaline lysis protocol and can be divided into the following steps:The bacteria are partially lysed, allowing the plasmid DNA to escape the cell wall into the supernatant. The larger E. coli chromosomal DNA is trapped in the cell wall. The lysate is cleared of cellular debris by filtration or centrifugation, and the plasmid DNA-containing fraction is loaded onto a pre-equilibrated column. Since the cleared lysate is applied to the column in a low-salt buffer, plasmid DNA binds to the macroporous anion-exchange material in the column. Cellular impurities are eluted from the column with a high-salt wash. Finally, the plasmid DNA is eluted from the column. Recovered DNA is precipitated from the eluate to remove salt and concentrate the plasmid.
Analysis Note
Sample:
E. coli culture that contains a high-copy number plasmid: 5 to 30 mL bacterial culture
E. coli culture that contains a low-copy number plasmid: 10 to 100 mL bacterial culture
Plasmid Size: The isolation procedure is suitable for all sizes of plasmid. Note: Lysates of larger constructs (up to 100 kb) should be cleared by filtration rather than centrifugation to avoid shearing the plasmid.
Time Required: 60 minutes (including filtration of the lysate)
Typical Yield:
High-copy number plasmid: 3 to 5 µg/mL culture
Low-copy number plasmid: 0.2 to 1 µg/mL culture
Product Purity: Isolated plasmid DNA is free of other bacterial components, including RNA.Other NotesIon Exchange Chromatography,
For life science research only. Not for use in diagnostic procedures.ПараметрыQuality Level 100, Торговая марка Roche packaging kit of for 20 isolations from 5 to 30 ml
Safety Informationpictograms GHS02,GHS05 signalword Danger hcodes H226 - H290 - H314 pcodes P210 - P280 - P303 + P361 + P353 - P304 + P340 + P310 - P305 + P351 + P338 + P310 - P370 + P378 RIDADR UN 3316 9 WGK Germany WGK 1 Flash Point F 100.4 °F Flash Point C 38 °C -
10127647001
Glucose-6-phosphate 10127647001
Empirical Formula (Hill Notation): C6H11Na2O9P · xH2O CAS Number: 3671-99-6 Molecular Weight: 304.10 (anhydrous basis) Beilstein/REAXYS Number: 5199009 MDL number: MFCD00150940 PubChem Substance ID: 329748950 General descriptionDisodium saltApplicationSubstrate for glucose-6-phosphatase.
Biochem/physiol Actions
In mammals, glucose-6-phosphate (G6P) is formed by a kinase acting on glucose. It plays several roles and has its fate determined accordingly. It can get converted back to glucose by a phosphatase enzyme. It can pass into glycogen, or enter into the energy-yielding Embden-Meyerhof path or into the 6-phosphogluconate pathway. Increased levels of blood glucose result in elevated levels of glucose 6-phosphate in liver, skeletal muscle, and adipose tissue. This elevated intracellular level of G6P activates glycogen synthase. G6P may also be involved in the negative regulation of phosphorylation of glycogen synthase via cyclic AMP-stimulated protein kinase.
Components
77% glucose-6-P (enzymatic), 13% sodium, 8% water
Quality
Contaminants: <0.2% glucose (enzymatic)
Formula variant
C6H11O9PNa2Other NotesFor life science research only. Not for use in diagnostic procedures.Параметрыmol wt (glucose-6-P: Mr = 260.2, glucose-6-P-Na2: Mr = 304.2) packaging pkg of 5 g Торговая марка Roche shipped in ambient SMILES string O.[Na+].[Na+].OC1O[C@H](COP([O-])([O-])=O)[C@@H](O)[C@H](O)[C@H]1O InChI 1S/C6H13O9P.2Na.H2O/c7-3-2(1-14-16(11,12)13)15-6(10)5(9)4(3)8;;;/h2-10H,1H2,(H2,11,12,13);;;1H2/q;2*+1;/p-2/t2-,3-,4+,5-,6?;;;/m1.../s1 InChI key UUWJZXLTPORJKW-WYFATIGWSA-L
Safety InformationRIDADR NONH for all modes of transport WGK Germany WGK 1 Flash Point F does not flash Flash Point C does not flash -
10165875001
Glucose-6-Phosphate Dehydrogenase (G6P-DH) 10165875001
Enzyme Commission (EC) Number: 1.1.1.49 ( BRENDA | IUBMB ) General descriptionGlucose-6-Phosphate Dehydrogenase (G6PD) is a ubiquitous enzyme which acts as a catalyst in producing pentose. It also produces NADPH for various biosynthetic and detoxification reactions. Sometimes G6PD provides an alternative to main glycolytic pathway for glucose utilization. It also has its application as DNA markers on X-chromosome.1
Specificity
Specificity: At pH 7.8, 25 °C: G6P-DH from Leuconostoc (LG6PDH) is highly specific for D-glucose-6-phosphate (Km = 36 µM, NADP as coenzyme; 64 µM, NAD as coenzyme), but will use either NADP (Km = 7.4 µM; relative rate 1.0) or NAD (Km = 115 µM; relative rate 1.8) as coenzyme.
LG6P-DH does not react with fructose-6-phosphate, fructose-1,6-biphosphate, glucose-1-phosphate or ribose-1-phosphate. LG6P-DG will oxidize 2-deoxy-glucose-6-phosphate with NADP, but not with NAD as coenzyme. There is a slow reaction with D-glucose.
Heat inactivation: The ammonium sulfate suspension is not inactivated when heated to temperatures 50 °C for 10 minutes. At temperatures > 60 °C the enzyme is rapidly inactivated.ApplicationComponent of cofactor recycling systems for NADPH.
It was used in enzymatic determination of C6 phosphorylation of glycogen.
Quality
Contaminants: <0.001% CK, <0.01% GR and PGI each, <0.02% “NADH oxidase”, <0.05% HK, <0.001% 6-PGDH
Unit Definition
Unit Conversion: One unit (U) [+25 °C] 1.18 U [+30 °C] 1.45 U [+37 °C], all with NAD as coenzyme
Unit Definition: One unit (U) LG6P-DH oxidizes 1 mol of glucose-6-phosphate and reduces 1 mol of NAD in 1 minute at +25 °C and pH 7.8.
Physical form
Solution of 1,000 U in 1 ml 3.2 M ammonium sulfate, pH approximately 6
Preparation Note
Activator: HCO3- ( 0.3 M) activates slightly.
Analysis Note
Absorbance of purified enzyme: 1.15 (1 mg enzyme/ml, 280.5 nm)Other NotesFor life science research only. Not for use in diagnostic procedures.ПараметрыQuality Level 100, form solution, suspension specific activity ~550 units/mg protein (At 25 °C (650 U/mg at 30 °C) with glucose-6-P and NAD as the substrates.) packaging pkg of 1 mL (1,000 U) Торговая марка Roche optimum pH 7.0-8.5(maximal activity at 7.8) 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 -
10127655001
Glucose-6-Phosphate Dehydrogenase (G6P-DH) 10127655001
Enzyme Commission (EC) Number: 1.1.1.49 ( BRENDA | IUBMB ) General descriptionGlucose-6-phosphate dehydrogenase (G6P DH) has an active dimeric form, whereas its monomeric and tetrameric forms are inactive. At low NADP+ and NADPH levels and under normal conditions, the enzyme exists as equilibrium of tetramers and dimers, whereas at high NADP+ and NADPH levels the equilibrium shifts to inactive tetramers.ApplicationComponent of cofactor recycling systems for NADPH.
Biochem/physiol Actions
Glucose-6-phosphate dehydrogenase (G6P-DH) is involved in the pentose phosphate pathway. It is responsible for catalysing the oxidation of glucose-6-phosphate (G6P) to gluconolactone-6-phosphate, with NADP+ simultaneously accepting hydrogen to form NADPH, thereby maintaining NADPH levels in cells.
Specifications
Contaminants: <0.001% CK, <0.01% GR, HK, 6-PGDH, and PGluM each, <0.002% PGI each, <0.002% PGI
Approximately 350U/mg at +25°C with glucose-6-phosphate as the substrate.
Physical form
Suspension in 3.2 M ammonium sulfate solution, pH approximately 6
Preparation Note
Activator: Mg2+ (5 to 10 mM)Other NotesFor life science research only. Not for use in diagnostic procedures.ПараметрыQuality Level 100, form suspension specific activity ~350 units/mg protein (At 25 °C with glucose-6-P as the substrate.) packaging pkg of 1 mL (5 mg/ml) Торговая марка Roche optimum pH 9.2 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 -
10197734001
Glutamate Dehydrogenase (GlDH) 10197734001
Enzyme Commission (EC) Number: 1.4.1.3 ( BRENDA | IUBMB ) General descriptionGDH (Glutamate dehydrogenase) is a hexamer of 449 residues and is located in the mitochondria. It acts as a branch-point enzyme between amino acid oxidation and urea production. L-glutamate:NAD(P)+ oxidoreductase is involved in deamination.ApplicationGlutamate dehydrogenase has been used to measure residual ammonium by enzymatic analysis during fermentation by wine yeast. It used as a component of cofactor recycling systems for NAD(P) and NAD(P)H.
Quality
Contaminants: <0.005% ADH, LDH, and MDH, each
Sequence
The smallest active structure of GIDH (310,000 to 350,000 ) is a hexamer subunit of 55,400 D. This hexamer reversibly self-assembles forming a polymer with up to eight hexamers (2,200 kD).Other NotesFor life science research only. Not for use in diagnostic procedures.ПараметрыQuality Level 100, form lyophilized specific activity 10 U/mg (Approximately 10 U/mg lyophilizate (120 U/mg enzyme protein) at +25°C with 2-oxoglutarate as the substrate, and ADP as the activator.), ~10 units/mg protein (at 25 °C with 2-oxoglutarate as the substrate, and ADP as the activator.) packaging pkg of 3,000 U Торговая марка Roche optimum pH 8 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 -
Glutamate-Oxaloacetate Transaminase (GOT)
Enzyme Commission (EC) Number: 2.6.1.1 ( BRENDA | IUBMB )
Кат. номер 10105546001 10737046001 10105554001 General descriptionGlutamate-Oxaloacetate Transaminase (GOT) is a transaminase enzyme which catalyzes amino group of an amino acid with keto group of keto acid.
L-aspartate:2-oxoglutarate aminotransferaseApplicationSynthesis of unnatural L-amino acids from α-keto acids.
Unit Definition
Unit Conversion: One unit (+25 °C, pH 7.4) ≈ 1.9 units (+37 °C, pH 7.4)
Physical form
Suspension in 3.2 M ammonium sulfate solution, 2.5 mM 2-oxoglutarate, 50 mM maleate, and 1 mM pyridoxal phosphate, pH approximately 6
Preparation Note
Activator: Pyridoxal phosphate, pyridoxamine phosphateOther NotesFor life science research only. Not for use in diagnostic procedures.ПараметрыQuality Level 100, form suspension specific activity ~200 units/mg protein (At 25 °C (380 U/mg at 37 °C) with L-aspartate and 2-oxoglutarate as the substrates) packaging pkg of 1 mL (10105546001 [2 mg]), pkg of 1 mL (10105554001 [10 mg]), pkg of 2.5 mL (10737046001 [25 mg]) Торговая марка Roche optimum pH 6.5-8.5 shipped in wet ice storage temp. 2-8°C
Safety InformationRIDADR NONH for all modes of transport -
Glutamate-Pyruvate Transaminase (GPT)
Enzyme Commission (EC) Number: 2.6.1.2 ( BRENDA | IUBMB )
Кат. номер 10737127001 10105589001 General descriptionGlutamate-pyruvate transaminase is a blood enzyme. It metabolizes glutamate in blood resulting in low extracellular levels of brain glutamate.ApplicationSynthesis of unnatural L-amino acids from α-keto acids.
Quality
Contaminants: < 0.01% GIDH, LDH, and MDH each, < 0.03% GOT
Unit Definition
Unit Conversion: 1 U [+25 °C, pH 7.5] ≈ 1.75 U [+37 °C, pH 7.5]
Physical form
Suspension in 3.2 M ammonium sulfate solution, pH approximately 6
Preparation Note
Activator: Pyridoxalphosphate
Storage conditions (working solution): The enzyme is stable when dissolved in human serum and frozen in aliquots. For best results, centrifuge the enzyme and decant the supernatant before dissolving the enzyme in human serum to avoid high ammonium sulfate concentrations in the solution.Other NotesFor life science research only. Not for use in diagnostic procedures.ПараметрыQuality Level 100, form suspension specific activity ≥80 units/mg protein (At 25 °C with L-alanine and 2-oxoglutarate as the substrates.) packaging pkg of 1 mL (10105589001 [10 mg]), pkg of 2.5 mL (10737127001 [25 mg]) Торговая марка Roche optimum pH ~8.0 shipped in wet ice storage temp. 2-8°C
Safety InformationRIDADR NONH for all modes of transport -
10901393001
Glycogen 10901393001
ApplicationThis preparation is used as a carrier for the precipitation of nucleic acids (DNA or RNA). As an inert material it may replace tRNAs or sonicated DNAs.
20 µg glycogen (1 µl solution) allow to precipitate pg-amounts of DNA or RNA from a volume of 1 ml.
In a typical experiment 5 pg [3H]-labeled calf thymus DNA were dissolved in 500 µl 10 mM Tris-HCl, pH 8.0; 1 mM EDTA; 0.4 M LiCl. 1µl glycogen solution (20 µg glycogen) as carrier was added and then precipitated with 1.2 ml ethanol at -15 to -25 °C and stored for 3 hours at -15 to -25 °C. After centrifugation (10 minutes at 12 000 x g) the total radioactivity was found in the precipitate. Without addition of glycogen no precipitation of DNA occured.
Features and Benefits
Glycogen, in special quality for molecular biology, is an inert carrier in nucleic acid preparations.
Contents
Aqueous solution, 20 mg/ml
Quality
Tested for absence of endonucleases, nicking activity, exonucleases, RNases, nucleic acids, and proteases according to the current Quality Control procedures.Other NotesFor life science research only. Not for use in diagnostic procedures.ПараметрыQuality Level 100, form solution packaging pkg of 1 mL (20 mg) Торговая марка Roche 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 -
10220647001
GTP--S 10220647001
General descriptionGuanosine 5-O-(3-thiotriphosphate), tetralithium salt (GTP--S) is a non-hydrolysable analog of GTP. It is an inhibitor of phosphodiesterase in photoreceptors.ApplicationGTP--S has been used:- in G-Protein activation assay to assess the functionality of protease-activated receptor 4 (PAR4)
- in fluorimetric guanine nucleotide exchange assay of G protein alpha subunits (G)
- to monitor the Rac family small guanosine triphosphatase (GTPase) 2 (Rac2)-stimulated activity of phospholipase C2 mutants
Biochem/physiol Actions
Guanosine 5-O-(3-thiotriphosphate) (GTP--S) activates guanine-nucleotide-binding proteins and is slowly hydrolyzed enzymatically. It inhibits guanosine triphosphatase (GTPases) more potently than guanosine triphosphate (GTP),
Storage and Stability
Store powder dry at -15 to -25 °C; aqueous solutions with pH approx. 7 should be stable for six months at -15 to -25 °C.
Analysis Note
Absorption: Content is measured in aqueous solution by absorption at 254 nm.
Conditions:- approx. 10 mg/ml double-dist. water
- dilution: 9.8 ml double-dist. water + 0.2 ml sample.
Other NotesFor life science research only. Not for use in diagnostic procedures.Параметрыdescription C10H12N5O13P3SLi4 Quality Level 100, assay 1% (GMP, HPLC), 12% (GDP, HPLC), 2% (GTP, HPLC) mol wt Mr 539.2 (GTP--S), Mr 563.0 (GTP--S-Li4) packaging pkg of 10 mg Торговая марка Roche storage condition protect from light shipped in dry ice storage temp. 20°C (15°C to 25°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 -
11685929001
Guanidine thiocyanate 11685929001
Empirical Formula (Hill Notation): CH5N3 · CHNS Linear Formula: NH2C(=NH)NH2 · HSCN CAS Number: 593-84-0 Molecular Weight: 118.16 Beilstein/REAXYS Number: 3563461 MDL number: MFCD00013027 PubChem Substance ID: 329749530 General descriptionGuanidine thiocyanate is chaotropic salts which lyse cells and along with nuclease binding matrix it is also seen to inhibit nuclease activity during extraction of DNA. It works as a strong RNase inhibitor in extraction buffers.ApplicationSuitable as denaturing agent for proteins. Guanidine thiocyanate is a stronger denaturant than guanidine hydrochloride.
Chaotropic agent and strong denaturant; solubilizes cells.
Specifications
Absorbance: A300 and A410 0.10 (6 M in H2O) each
Formula: CH5N3 x HSCN
Physical form
CrystalsOther NotesFor life science research only. Not for use in diagnostic procedures.Параметрыmol wt 118.16 packaging pkg of 500 g Торговая марка Roche mp 120-122 °C (lit.) absorption ≤0.10 at 300 and 410 nm in water at 6 M shipped in ambient storage temp. 20-25°C SMILES string SC#N.NC(N)=N InChI 1S/CH5N3.CHNS/c2-1(3)4;2-1-3/h(H5,2,3,4);3H InChI key ZJYYHGLJYGJLLN-UHFFFAOYSA-N
Safety Informationpictograms GHS05,GHS07 signalword Danger hcodes H302 + H312 + H332 - H314 - H412 pcodes P260 - P273 - P280 - P303 + P361 + P353 - P304 + P340 + P310 - P305 + P351 + P338 + P310 Supp Hazards EUH032 RIDADR NONH for all modes of transport WGK Germany WGK 2 Flash Point F Not applicable Flash Point C Not applicable -
11666975001
HA Peptide 11666975001
General descriptionHA (hemagglutinin) is a surface glycoprotein, expressed by influenza virus.ApplicationHA Peptide has been used in the following :- Protein purification from Baculovirus infected cells.
- Affinity chromatography.
- Chromatin immunoprecipitation.
Biochem/physiol Actions
HA (hemagglutinin) protein is responsible for the infectivity of human influenza by inducing the production of neutralizing antibodies. HA mediates viral attachment to the surface of oligosaccharide receptors (at sialic acid terminal). HA contains conserved cross-reactive antigenic determinants, which is predicted to stimulate cross-protective antibodies.
Quality- Purity: =95% (HPLC)
- Function test: Competitive ELISA, using Anti-HA-coated microplate to measure free HA Peptide inhibition of HA-peroxidase binding with detection by ABTS.
Reconstitution
Add 5 ml TBS to get a final concentration of 1 mg/ml.
Rehydrate for at least 10 minutes.Other NotesFor life science research only. Not for use in diagnostic procedures.Параметрыassay 95% (HPLC) form lyophilized packaging pkg of 5 mg Торговая марка Roche shipped in dry ice storage temp. −20°C
Safety InformationRIDADR NONH for all modes of transport WGK Germany WGK 1 Flash Point F Not applicable Flash Point C Not applicable -
11426362001
Hexokinase (HK) 11426362001
Enzyme Commission (EC) Number: 2.7.1.1 ( BRENDA | IUBMB ) General descriptionATP:D-hexose 6-phosphotransferase
Hexokinase exists in three isozymes, namely type I, type II and type III that differ in subcellular localization, catalytic and regulatory properties. The type I isozyme serves a catabolic function facilitating the entry of glucose into glycolytic metabolism for energy production. The type II and type III enzymes perform anabolic functions, such as generating G6P for glycogen synthesis or metabolism through the phosphogluconate pathway for lipid synthesis. Type I and type II isozymes have been found to be bound to mitochondria. The type III isozyme has been found to have perinuclear localization in several cell types.
Hexokinase consists of 486 amino acids and has a molecular weight of 57,000 D (SDS-PAGE) in citrate/phosphate buffer. It may form dimers under other buffer conditions.ApplicationHexokinase (HK) has been used in enzyme assays and chemiluminescence assay. It has also been used to determine ATP concentration.
Use Hexokinase for the determination of D-glucose, D-fructose, and D-sorbitol in food or biological research samples. Hexokinase may also be used for the assay of other saccharides, which are convertible to glucose or fructose, and is, therefore, useful in the assay of many glycosides.
Hexokinase has been used in the enzymatic detection of nitric oxide and in the measurement of ATP by luminescence method.
Biochem/physiol Actions
Hexokinase catalyzes the conversion of hexoses to hexose-6-phosphate in an ATP-dependent manner. It is also called as the glycolytic enzyme as it catalyzes the first step in glucose metabolism with the formation of glucose-6-phosphate (G6P). In eukaryotes, it functions as a glucose sensor.
Quality
Contaminants: <0.002% PGI, <0.0005% G6P-DH and GR, each, <0.001% CK, MK, 6-PGDH, and ADH, each, <0.05% GIDH and invertase (-fructosidase), each, <30 µg/ml glucose (enzymatically)
Unit Definition
Unit Assay: For measuring hexokinase activity 0.22 M glucose in 0.1 M triethanolamine buffer, pH 7.6, 6.5 mM MgCI2, 2.7 mM ATP, 0.83 mM NADP is incubated in the presence of 1.7 U glucose-6-phosphate dehydrogenase with an appropriate amount of hexokinase (10–20 mU) at +25 °C (total volume: 3.0 ml). The enzyme activity is calculated from the increase of absorbance at 340 nm ( = 6.3 [I x mmol/l-1 x cm-1]) or 365 nm ( = 3.5 [I x mmol/l-1 x cm-1]).
Physical form
Suspension in 3.2 M ammonium sulfate solution, pH approximately 6.5
Preparation Note
Activator: Hexokinase requires Mg2+ ions (Km = 2.6 mM) for activity. Catecholamines too increase activity.
Stabilizers: ThiolsOther NotesFor life science research only. Not for use in diagnostic procedures.Параметрыform suspension Quality Level 100, specific activity ~450 units/mg protein (At 25 °C and pH 7.6 with D-glucose and ATP as the substrates.) packaging pkg of 1 mL (1,500 U), pkg of Торговая марка Roche pH range 6.0 - 8.0 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 -
Hexokinase/Glucose-6-Phosphate Dehydrogenase (HK/G6P-DH)
Кат. номер 10737275001 10127825001 General descriptionHexokinase is responsible for catalyzing the first step of glycolysis. It results in the formation of glucose-6-phosphate and ADP from glucose and ATP. Glucose-6-phosphate dehydrogenase is a crucial enzyme in the pentose phosphate pathway. It is responsible for the conversion of glucose-6-phosphate and NADP+ (nicotinamide adenine dinucleotide phosphate) to 6-phosphogluconolactone and NADPH.ApplicationHexokinase/Glucose-6-phosphate dehydrogenase has been used for the determination of plasma glucose levels.
Components
EC 2.7.1.1 & 1.1.1.49
Unit Definition
One unit (U) HK will phosphorylate 1 µmol of D-glucose in one minute at +25 °C and pH 7.6.
One unit (U) G6P-DH will oxidize 1 µmol of glucose-6-phosphate in one minute at +25 °C and pH 7.6.
The coupled assay produces 1 µmol of NADH per µmol of D-glucose phosphorylated.
Volume Activity: 340 U hexokinase/ml at +25 °C with glucose and ATP as the substrates. 170 U glucose-6-phosphate dehydrogenase/ml at +25 °C with glucose-6-phosphate as the substrate
Physical form
Suspension in 3.2 M ammonium sulfate solution, pH approximately 6. Prepared by mixing hexokinase with G6P-DH. HK:G6P-DH approximately 2:1 with regard to protein content.Other NotesFor life science research only. Not for use in diagnostic prodcedures.ПараметрыQuality Level 100, form suspension packaging pkg of 10 mL (10737275001 [30 mg]), pkg of 5 mL (10127825001 [15 mg]) Торговая марка Roche shipped in wet ice storage temp. 2-8°C
Safety InformationRIDADR NONH for all modes of transport -
11585878001
hGH ELISA 11585878001
General descriptionThe hGH assay system differs from most of the other commonly used reporter proteins (CAT, -Gal, or luciferase) in one important respect: hGH is a secreted protein, and is measured using samples of culture-medium supernatants thus (i) avoiding the necessity to lyse cells, (ii) allowing continuous monitoring of transient expression kinetics, and (iii) allowing use of the cells for other purposes (e.g., RNA isolation). Additionally, hGH is a mammalian gene that may contribute to the apparent high stability of the hGH mRNA in most mammalian cells.
Using the standard isotopic protocol, secretion of hGH protein into the culture-medium supernatant can be monitored approximately 24hours post-transfection, depending on the cell type and the sensitivity of the assay used. Usually, hGH is quantified by an immunoradiometric assay (IRMA), resulting in a linear range of approximately 0.1 to 50ng hGH/ml.
Human growth hormone (hGH) is specifically detected by the hGH ELISA. It does not cross-react with rat growth hormone. Cross-reactivity with the human homologues TSH, FSH, and LH has not been tested. The hGH protein from E. coli, included in the kit for the purpose of compiling a standard calibration curve, is provided with lot-specific content data as determined by immunoassay.
hGH Standards
The hGH protein from E. coli, included in the kit for the purpose of compiling a standard calibration curve, is provided with lot-specific content data as determined by immunoassay.
Specificity
The hGH ELISA specifically detecs human growth hormone (hGH). It does not cross-react with rat growth hormone.ApplicationFor research use only. Not for use in diagnostic procedures.
The hGH ELISA is used to quantitatively measure the expression level of hGH released into cell culture supernatant of eukaryotic cells transfected with a plasmid containing a reporter gene that encodes for hGH.
Features and Benefits- Standardized: Allows direct comparison of data from different sets of experiments, even when kits from different production lots are used
- More sensitive: The hGH ELISA is approximately 20-times more sensitive than isotopic hGH assays
- More specific: The hGH ELISA shows no crossreactivity with rat growth hormone
- Faster: Measures hGH expression as early as 18hours post-transfection and requires only approximately 4 hours from start to finish
- Easy to perform: Follows a standard ELISA protocol
Packaging
1 kit containing 9 components.
Specifications
Assay time: Approximately 4 hours
Sample material: Culture supernatant
Sensitivity: 5pg/ml (1pg/well)
Standards: The hGH protein from E. coli, included in the kit for the purpose of compiling a standard calibration curve, is provided with lot-specific content data as determined by immunoassay.
Principle
The hGH ELISA is based on the sandwich ELISA principle. Antibodies to hGH (anti-hGH) are prebound to the surface of the microplate. The culture supernatant, which contains secreted hGH, is added to the wells of the microplate modules. All hGH contained in the medium binds to the anti-hGH bound to the microplate surface. Next, a digoxigenin-labeled antibody to hGH (anti-hGH-DIG) is added, then bound to hGH. In the following step, an antibody to digoxigenin conjugated to peroxidase (anti-DIG-POD) is added, then bound to digoxigenin. In the final step, the peroxidase substrate ABTS is added. The peroxidase enzyme catalyzes the cleavage of the substrate, yielding a colored reaction product. The absorbance of the sample is determined using an ELISA reader, and is directly correlated to the level of hGH present in the medium supernatant. The sensitivity of the assay can be enhanced using the peroxidase substrate (ABTS) with substrate enhancer.
Preparation Note
Working solution: Solution 1 hGH Stock Solution (bottle 1)
Reconstitute the lyophilizate in 0.5 ml double distilled water. The resulting concentration is calculated using the lot-specific information, given on the label (final conc. approx 10 ng/ml). This solution is used for the preparation of hGH standards for establishing a hGH calibration curve (see 7.2: preparation of hGH standards).
Solution 2 Anti-hGH-DIG (bottle 2)
Reconstitute the lyophilizate in 0.5 ml double distilled water (final conc. 100 µg/ml).
Solution 2a Anti-hGH-DIG, working dilution
To prepare anti-hGH-DIG working dilution, dilute the reconstituted anti-hGH-DIG solution (100 g/ml) with sample buffer (solution 7) to a final conc. of 1 µg/ml (e.g., 100 µl of reconstituted anti-hGH-DIG solution 9.9 ml of solution 7 for 50 wells).The working dilution should be prepared freshly before use and should not be stored.
Solution 3 Anti-DIG-POD (bottle 3)
Reconstitute the lyophilizate in 0.5 ml double distilledwater (final conc. 20 U/ml). Do not add sodiumazide!
Solution 3a Anti-DIG-POD, working dilution
To prepare anti-DIG-POD, working dilution, dilute the reconstituted anti-DIG-POD solution (20 U/ml) with sample buffer (solution 7) to a final conc. of 200 mU/ml (e.g., 100 µl of reconstituted anti-DIG-POD solution 9.9 ml of solution 7 for 50 wells). The working dilution should be prepared freshly before use and should not be stored.
Solution 4 POD Substrate (bottle 4)
Ready-to-use solution.
Solution 5 POD Substrate containing Substrate Enhancer (bottles 4 and 5)
If a low hGH concentration in the sample is expected, add 1 mg of substrate enhancer (bottle 5) per ml of solution 4 and mix by stirring for 30 min at 15 to 25 °C. The solution is stable for only 2 hours and should therefore be prepared immediately before use. Use the substrate enhancer only if the hGH concentration is low!
Solution 6 Washing Buffer, 1x (bottle 6)
To prepare a ready-to-use washing buffer, mix 1 part 10x washing buffer concentrate (bottle 6) with 9 parts of double distilled water.
Solution 7 Sample Buffer (bottle 7)
The solution is ready-to-use. Mix thoroughly before use. Do not add sodium azide.
Storage conditions (working solution): Solution 1 hGH Stock Solution (bottle 1)
The reconstitutedsolution is stable for 2 months at 2 to 8 °C. For long term storage, store in aliquots at or below -15 to -25 °C.
Solution 2 Anti-hGH-DIG (bottle 2)
The reconstituted solution is stable for two months at 2 to 8 °C. For long term storage, store in aliquots at or below -15 to -25 °C.
Solution 2a Anti-hGH-DIG, working dilution
The working dilution should be prepared freshly before use and should not be stored.
Solution 3 Anti-DIG-POD (bottle 3)
The reconstituted solution is stable for 6 months at 2 to 8 °C. Do not freeze!
Solution 3a Anti-DIG-POD, working dilution
The working dilution should be prepared freshly before use and should not be stored.
Solution 4 POD Substrate (bottle 4)
The solution is stable until the expiration date given on the label if stored at 2 to 8 °C.
POD Substrate containing Substrate Enhancer (bottles 4 and 5)
The solution is stable for only 2 hours and should therefore be prepared immediately before use.
Solution 6 Washing Buffer, 1x (bottle 6)
The reconstituted solution is stable for 6 months at 2 to 8 °C.
Solution 7 Sample Buffer (bottle 7)
After opening the bottle we recommend storing the solution in aliquots at -15 to -25 °C, since it does not contain a preservative agent.
Analysis Note
No cross-reaction with rat growth hormone. Cross reactivity with the human homologues TSH, FSH, and LH has not been tested.Other NotesFor life science research only. Not for use in diagnostic procedures.Параметры