The molecular weight for this product is not determined. However, other sources report a molecular weight of 24,000 daltons for Diaphorase from Clostridium.
D2197
Diaphorase from Clostridium kluyveri
recombinant, expressed in E. coli, Animal-component free
Synonym(s):
Diaphorase, Lipoamide Dehydrogenase, Lipoyl Dehydrogenase
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Product Name
Diaphorase from Clostridium kluyveri, recombinant, expressed in E. coli, Animal-component free
form
solid
recombinant
expressed in E. coli
specific activity
>=30 units/mg protein (biuret)
storage temp.
−20°C
Quality Level
1 of 4
This Item | D5540 | 37294 | 10171832001 |
|---|---|---|---|
| specific activity >=30 units/mg protein (biuret) | specific activity 3.0-20.0 units/mg protein (biuret) | specific activity ≥6 U/mg protein | specific activity ~20 units/mg protein (At 25 °C with acetaldehyde as the substrate.) |
| recombinant expressed in E. coli | recombinant - | recombinant - | recombinant - |
| form solid | form lyophilized powder | form ammonium sulfate suspension | form lyophilized |
| storage temp. −20°C | storage temp. −20°C | storage temp. 2-8°C | storage temp. 2-8°C |
| Quality Level 200 | Quality Level 200 | Quality Level 100 | Quality Level 100 |
Application
Diaphorase from Clostridium kluyveri has been used:
- in lactate dehydrogenase assay in gill cells.[1]
- impregnated on the strip for formate dehydrogenase assay.[2]
- in various assays to oxidate β-NADH or β-NADPH in the presence of an electron acceptor.
- for semiquinone generation during benzoquinone reduction.[3]
- for evaluation of glucose uptake in cultured muscle cells.[4]
- in lactate dehydrogenase cytotoxicity assay.[5]
Biochem/physiol Actions
Diaphorase catalyzes the oxidation of either β-NADH or β-NADPH in the presence of an electron acceptor such as methylene blue or 2,6-dichlorophenolindophenol. Diaphorases that are specific for either β-NADH or β-NADPH are known. The pig heart enzyme of Straub seems to have native diaphorase (β-NADH specific) as well as lipoic and lipoamide dehydrogenase activities.
General description
Research area: Cancer
Other Notes
One unit of diaphorase will oxidize 1.0 micromole of beta-NADH per minute at pH 7.5 at 25 degrees C, with the corresponding reduction of the appropriate electron acceptor.
Physical form
Supplied as a lyophilized powder containing trehalose.
Storage Class
11 - Combustible Solids
wgk
WGK 3
flash_point_f
Not applicable
flash_point_c
Not applicable
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What is the molecular weight of this diaphorase?
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How to prepare stock solution for Diaphorase in order to use 2 units in reaction
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For assay purposes, this product is reconstituted 1 mg/ml in cold 200 mM Tris-HCl buffer containing 294 mM Potassium Chloride, 0.54 mM Riboflavin 5'-monophosphate, and 0.025% (w/v) BSA, pH 7.5. The minimum activity of this enzyme is 30 u/mg protein with a minimum protein content of 50%. At the minimum activity and minimum protein content, 2 units of 1 mg/mL stock solution would have a volume of 133 uL. Please see the link below to review a sample or lot-specific Certificate:
https://www.sigmaaldrich.com/product/sigma/d2197#product-documentationHelpful?
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