Description
Description: This assay employs a two-site sandwich ELISA to quantitate CASP3 in samples. An antibody specific for CASP3 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any CASP3 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for CASP3 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 CASP3 bound in the initial step. The color development is stopped and the intensity of the color is measured.
Overview: Caspase-3 (CPP-32, SCA-1) is a critical executioner of apoptosis, as it is either partially or totally responsible for the proteolytic cleavage of many key proteins such as the nuclear enzyme poly (ADP-ribose) polymerase (PARP) (1). Activation of caspase-3 requires proteolytic processing of its inactive zymogen into activated p17 and p12 fragments. Cleavage of caspase-3 requires aspartic acid at the P1 position (2).a protein that is a member of the cysteine-aspartic acid protease (caspase) family. Sequential activation of caspases plays a central role in the execution-phase of cell apoptosis. Caspases exist as inactive proenzymes that undergo proteolytic processing at conserved aspartic residues to produce two subunits, large and small, that dimerize to form the active enzyme