Amplite(TM)Fluorimetric Alkaline Phosphatase Assay Kit *Red Fluorescence*

Alkaline phosphatase is widely used in various biological assays (in particular, immunoassays) and ELISA-based diagnostics. Our Amplite(TM) Fluorimetric Alkaline Phosphatase Assay kit uses our SunRed(TM)-based substrate. The weakly fluorescent SunRed(TM) phosphate is sensitive to phosphatase-induced hydrolysis, giving the SunRed(TM) fluorophore that possesses intense red fluorescence. Upon phosphatase-induced hydrolysis, the SunRed(TM) phosphate solution has its absorption blue-shifted more than 100 nm. The maximum absorption of SunRed(TM) fluorophore at 633 nm makes this substrate an ideal NIR probe that can be readily detected with many fluorescence instrument systems often equipped with Cy5 settings. Based on the near infrared fluorescence of SunRed(TM) fluorophore, the signal can be easily read by a fluorescence microplate reader at Ex/Em = 630/660 nm. The kit has been used for the high throughput screening of protein phosphatase inhibitors due to its low interference from biological sample. It can be performed in a convenient 96-well or 384-well microtiter-plate format and easily adapted to automation without a separation step.