NADPH Regenerating System Solution B
| Catalog Number: 451200 (Old B200) |
Storage Conditions: Store at -20°C |
| Lot Number: xx |
Date Released: Year Month |
| Package Size: 1 ml |
|
| NADP+ Reductase Activity:
xx µmoles/min/ml |
| Solution B Components: 40 U/ml Glucose-6-phosphate
dehydrogenase in 5 mM sodium citrate |
xx = Actual values can be found on
the data sheet accompanying each shipped product.
Description: NADPH is a necessary cofactor in many
xenobiotic metabolism reactions. NADPH is required for the measurement
of oxidase activity catalyzed by P450s, FMOs, NADPH-P450 reductase, and
many other oxidase enzymes. A common source of NADPH in an oxidase enzyme
assay is an NADPH regenerating system which generates NADPH in situ using
an enzymatic reaction. For example, glucose-6-phosphate dehydrogenase
(G6PDH) will convert NADP+ to NADPH in the presence of the substrate glucose-6-phosphate
(Glc-6-PO4).
The BD Gentest NADPH regenerating system consists of two reagents, Solution
A (NADP+ and Glc-6-PO4) and Solution B (G6PDH). Each reagent
is sold separately. Combined, these two reagents form an NADPH regenerating
system that can be used for all NADPH requiring oxidase assays (cDNA-expressed
enzymes and liver fractions).
General Use: For a typical oxidase activity assay, the recommended
concentrations for the components of an NADPH regeneration system are;
1.3 mM NADP+, 3.3 mM glucose-6-phosphate, 0.4 U/ml glucose-6-phosphate
dehydrogenase, and 3.3 mM magnesium chloride. If used at these concentrations,
Solutions A and B in the BD Gentest NADPH Regenerating System are 20X
and 100X respectively.
At least 200-400 enzyme assays can be performed using one vial each of
Solution A and B. The total number of assays that can be performed is
dependent on a researcher's experimental design.
Stability: Under investigation.
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