Detection Method:Sandwich
Test principle:This assay employs a two-site sandwich ELISA to quantitate PIK3AP1 in samples. An antibody specific for PIK3AP1 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyPIK3AP1 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for PIK3AP1 is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of PIK3AP1 bound in the initial step. The color development is stopped and the intensity of the color is measured.
Product Overview:Using an affinity purification strategy with the N-terminal SH2 domain of the phosphoinositide 3-kinase (PI3K) p85 subunit and anti-phosphotyrosine antibody, followed by screening a chicken DT40 B-cell cDNA library and a mouse spleen cDNA library, Okada et al. (2000) isolated cDNAs encoding chicken and mouse Pik3ap1, which they called Bcap. They also identified Bcap splice variants. Northern blot analysis of mouse tissues detected expression predominantly in spleen, with lower levels in thymus, liver, and lung. Expression was also detected in mouse macrophage and B-cell lines, but not in T-cell, plasma cell, or mast cell lines.Using mouse and chicken B- cell lines, Okada et al. (2000) determined that Bcap is a protein tyrosine kinase substrate that couples B-cell receptor to PI3K activation.
