U-Blot® Oxidated Afamin and Total Afamin ELISA Kit

The Oxidated Afamin 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 Afamin antibody has been coated onto a 96-well plate. Biotinylated samples are pipetted into the wells, and Afamin 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 Afamin. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of oxidated Afamin bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.This assay enables quick and easy quantification of target proteins and allows high-throughput screening of numerous different lysates without the need for traditional Western Blot analysis. It features minimal hands-on operation time, simple workflow, and completely non-radioactive detection system, ensuring safe and convenient experimental operation.