NADPH Regenerating System Solution A
| Catalog Number: 451220 (Old B220) |
Storage Conditions: Store at -20°C |
| Lot Number: xx |
Date Released: Year Month |
| Package Size: 5 ml |
| NADP+ Reductase Activity: xx
µmoles/min/ml |
| Solution A Components: 26.1 mM NADP+,
66 mM Glucose-6-phosphate, and 66 mM MgCl2 in H2O |
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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