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

Ex: 422 nm. Em: 466 nm. Nicotinamide adenine dinucleotide (NADH) and its oxidized form (NAD) are essential cofactors for many enzyme reactions found in living cells. 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 NADH or total NAD/NADH amount, but detection NAD generation in the presence of large excess amount of NADH has been quite challenging to date because NAD has its absorption peak at 259 nm and does not fluorescence, making the measurement unpractical. AmpliteT Fluorimetric NAD Assay Kit provides a sensitive and rapid detection of NAD. The kit directly measure NAD using Quest FluorT NAD reagent, our newly developed NAD sensor. The proprietary probe used in this kit reacts only with NAD to generate a product that fluorescence at Ex/Em = 420/480 nm, and has little response to NADH. This kit can detect as little as 30 nM NAD in a 100 µL assay volume, and monitor 0.3% NAD generation in the presence of excess amount of NADH. This assay can be performed in a convenient 96-well or 384-well microtiter-plate format and can be used in high-throughput screening.