In many protocols of glucose uptake assay kits or NAD(P)/NAD(P)H assay kits, NAD(P)+ is decomposed by boiling in approx. 80 degrees Celcius. I understood that NAD(P)+ is stable in acidic condition and reduced forms of those in alkali condition but, I'm wondering the composition of buffer in the above kits to decompose NAD(P)+. I made NADP+ solution in some buffers (pH was around 9) and measured the absorbance at 340 nm after boiling. But there was no decrease in the absorbance.
Nick Chacos
Remember that NADP does not absorb much at 340, only the reduced form does. To see if the NADP has decomposed, you could add a dehydrogenase and its substrate such as LDH/lactate and determine how much NADPH is generated.
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