BLNK (Phospho-Tyr96)Colorimetric Cell-Based ELISA Kit

A cell-based immunoassay from Assay Genie designed to detect phosphorylation of B-cell linker protein (BLNK) at tyrosine 96 in cultured cells. The kit is reactive with human and mouse samples and uses a colorimetric detection method read at 450 nm, with a stated dynamic range of "> 5000 Cells". It is intended for the adherent cell sample type and is classified under the Phospho sub-type for immunology research.

The assay operates on an indirect ELISA format in which BLNK (Phospho-Tyr96) is captured by phospho-specific primary antibodies and detected using HRP-conjugated secondary antibodies that catalyze a colorimetric reaction upon substrate addition. Presented as a lysate-free, high-throughput workflow, the kit can be used to measure relative levels of phosphorylated BLNK in cultured cells and to screen the effects of treatments, inhibitors (such as siRNA or chemicals), or activators on BLNK phosphorylation. Two normalization methods are provided: a monoclonal anti-human GAPDH antibody serves as an internal positive control, and a Crystal Violet whole-cell staining step may be used to normalize absorbance values to cell density.

BLNK is an adaptor protein that links B-cell receptor-associated kinases to downstream signaling pathways; phosphorylation of the protein on tyrosine residues by SYK and LYN is required for Ca(2+) mobilization and MAPK signaling. Target identifiers include UniProt: Q8WV28 and NCBI Gene ID: 29760.