NADH and NADPH Assay Kit (Colorimetric)

Product details

NADH and NADPH Assay Kit (ab186031) provides a convenient method for detecting NADH and NADPH.

How the NADH / NADPH assay works

Nicotinamide adenine dinucleotide (NAD+) and nicotinamide adenine dinucleotide phosphate (NADP+) are two important cofactors found in cells. NADH is the reduced form of NAD+. NAD forms NADP with the addition of a phosphate group to the 2' position of the adenyl nucleotide through an ester linkage. The traditional NAD/NADH and NADP/NADPH assays are based on monitoring the changes in NADH or NADPH absorption at 340 nm. The short UV wavelength of NAD/NADH and NADP/NADPH assays makes the traditional methods suffer low sensitivity and high interference.

In this NADH and NADPH assay method, the NADH/NADPH probe is a chromogenic sensor that has its maximum absorbance at 460 nm upon NADH/NADPH reduction. The absorbance maximum increases to ~ 635 nm if the enhancer is added to the assay system. The absorption of the NADH/NADPH probe is directly proportional to the concentration of NADH/NADPH in the solution. Note that this assay kit will measure both NADPH and NADH together, and cannot differentiate them.

The Colorimetric NADPH Assay Kit provides a sensitive assay to detect as little as 3 µM NADPH in a 100 µL assay volume.

NADH and NADPH assay protocol summary

- prepare NADPH Reaction Mixture

- add NADPH standards or test samples

- incubate at room temperature for 15-120 minutes

- monitor Absorbance at 460 nm

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