Amplite(TM) Fluorimetric NADP Assay Kit *Blue Fluorescence*

Ex: 422 nm. Em: 466 nm. Nicotinamide adenine dinucleotide (NAD+) and nicotinamide adenine dinucleotide phosphate (NADP+) are two important cofactors for many enzyme reactions found in living cells. NAD 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 requires NADPH as a reducing agent. In chloroplasts, NADP is an oxidizing agent important in the preliminary reactions of photosynthesis. The NADPH produced by photosynthesis is used as reducing power for the biosynthetic reactions in the Calvin cycle of photosynthesis. Quantifying the generation or consumption of these factors is an important method to monitor the enzyme-mediated reaction or screening the modulator or substrate of these enzyme reactions. There are several kits on the market to quantify NADPH or total NADP/NADPH amount, but detection NADP generation in the presence of large excess amount of NADPH has been quite challenging to date because NADP has its absorption peak at 260 nm and does not fluorescence, making the measurement unpractical. AmpliteT Fluorimetric NADP Assay Kit provides a sensitive and rapid detection of NADP. The kit directly measure NADP using Quest FluorT NADP reagent, our newly developed NADP sensor. The proprietary probe used in this kit reacts only with NADP to generate a product that fluorescence at Ex/Em = 420/480 nm, and has little response to NADPH. This kit can detect as little as 30 nM NADP in a 100 µL assay volume, and monitor 0.3% NADP generation in the presence of excess amount of NADPH. This assay can be performed in a convenient 96-well or 384-well microtiter-plate format and can be used in high-throughput screening.