NADPH is a critical cofactor supporting numerous biochemical reactions. In ADME-Tox studies, NAD(P)H regeneration is strongly recommended when using drug metabolizing enzymes (ex. Cytochrome P450 (CYP), Flavin-containing MonoOxygenases (FMO)), Recombinant CYPs (incl. bactosomes) or cellular fractions (Microsomes, S9). Currently, the most simple and cost-effective way to regenerate the NAD(P)H in situ and enzymatically is to use the commercially-available RapidStart™ NADPH Regenerating System (Xenotech-Sekisui).
RapidStart uses an enzymatic reaction that changes NADP to NADPH, which is then oxidized by CYPs back to NADP, and the cycle continues…
RapidStart™ is a versatile system that supports the long-term metabolism of test articles by drug metabolizing enzymes and sub-cellular fractions for in vitro drug assays. The system is based on Glucose-6-Phosphate DeHydrogenase (G6PDH) and Glucose-6-Phosphate (G6P), with reduction of NADP+ to NADPH. A most interesting advantage is that it’s extremely simple to use – just add water and use it!
NADPH RapidStart regenerating – versatile & simple ready-to-use system
RapidStart™ NADPH regenerating system is flexible since the end-user can conveniently modulate and adjust the NADPH concentration by simply adding water to it.
Simple protocol work flow: Add – Agitate – Activate!
- Determine the desired initial concentration using our RapidStart™ Dilution Concentration Tables (see below)
- Add the appropriate amount of high purity water in one single 0.3mL or 1.5 mL vial
- Vortex until pellets are fully dissolved & the NADPH regenerating system is activated and is ready for use!
What about you? Which NADPH method are you using in your ADME-Tox studies? Feel free to leave a message below to share your experience or to contact me for more information.