The RayBio^® Oxidated SIRT1 ELISA Kit uses the "biotin probes" method to biotin label carbonyl groups of oxidized proteins. In this method, the biotin probes first react with carbonyl groups of oxidized proteins to form unstable Schiff bases, which are then further reduced to more stable amines. The biotinylated proteins can then be detected by ELISA. An anti-pan SIRT1 antibody has been coated onto a 96-well plate. Biotinylated samples are pipetted into the wells, and SIRT1 present in a sample is bound to the wells by the immobilized antibody. The wells are washed, and HRP-conjugated streptavidin is used to detect oxidated SIRT1. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of oxidated SIRT1 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.

The entire kit may be stored at -20°C for up to 6 months from the date of shipment. Avoid repeated freeze-thaw cycles.

This assay can be developed for other species (mouse, rat, etc.) upon request. Contact techsupport@raybiotech.com for more information.