Detection Method:Sandwich
Test principle:This assay employs a two-site sandwich ELISA to quantitate APAF1 in samples. An antibody specific for APAF1 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyAPAF1 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for APAF1 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 APAF1 bound in the initial step. The color development is stopped and the intensity of the color is measured.
Product Overview:Apoptotic peptidase activating factor 1 is a proteincontains several copies of the WD40 repeat domain, a caspase recruitment domain (CARD), and an ATPase domain (NB-ARC). Upon binding cytochrome c and dATP, this protein forms an oligomeric apoptosome. The apoptosome binds and cleaves caspase 9 preproprotein, releasing its mature, activated form. The precise mechanism for this reaction is still debated though work published by Guy Salvesen suggests that the apoptosome may induce caspase 9 dimerization and subsequent autocatalysis. Activated caspase 9 stimulates the subsequent caspase cascade that commits the cell to apoptosis.Alternative splicing results in several transcript variants encoding different isoforms.
