Description
This assay employs a two-site sandwich ELISA to quantitate CASP14 in samples. An antibody specific for CASP14 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any CASP14 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for CASP14 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 CASP14 bound in the initial step. The color development is stopped and the intensity of the color is measured.
Overview: Caspase-14 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 which undergo proteolytic processing at conserved aspartic residues to produce two subunits, large and small, that dimerize to form the active enzyme. This caspase has been shown to be processed and activated by caspase 8 and caspase 10 in vitro, and by anti-Fas agonist antibody or TNF-related apoptosis inducing ligand in vivo. The expression and processing of this caspase may be involved in keratinocyte terminal differentiation, which is important for the formation of the skin barrier