Amplite(TM) Colorimetric NADP/NADPH Ratio Assay Kit

Ex: 635 nm. Nicotinamide adenine dinucleotide (NAD+) and nicotinamide adenine dinucleotide phosphate (NADP+) are two important cofactors found in cells. NADH is the reduced form of NAD+, and NAD+ is the oxidized form of NADH. It forms NADP with the addition of a phosphate group to the 2' position of the adenyl nucleotide through an ester linkage. NADP is used in anabolic biological reactions, such as fatty acid and nucleic acid synthesis, which require NADPH as a reducing agent. The traditional NAD/NADH and NADP/NADPH assays are done by monitoring of NADH or NADPH absorption at 340 nm. This method suffers low sensitivity and high interference since the assay is done in the UV range that requires expensive quartz microplate. Our AmpliteT NADP/NADPH Ratio Assay Kit provides a convenient method for sensitive detection of NADP, NADPH and their ratio. The NADPH probe is a chromogenic sensor that has its maximum absorbance at 635 nm upon NADH reduction. The absorbance increase at 635 nm is directly proportional to the concentration of NADPH in the solution. The NADPH probe can recognize NADPH in an enzyme-free reaction, and the signal can be easily read by an absorbance microplate reader at ~ 635 nm. The AmpliteT Colorimetric NADPH Assay Kit provides a sensitive assay to detect as little as 3 µM NADPH in a 100 µL assay volume. The assay can be performed in a convenient 96-well or 384-well microtiter-plate format.