Description
Description: This assay employs a two-site sandwich ELISA to quantitate ZDHHC5 in samples. An antibody specific for ZDHHC5 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any ZDHHC5 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for ZDHHC5 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 ZDHHC5 bound in the initial step. The color development is stopped and the intensity of the color is measured.
Overview: ZDHHC5 belongs to the DHHC palmitoyltransferase family. In molecular biology the DHHC domain is a protein domain that acts as an enzyme, which adds a palmitoyl chemical group to proteins in order to anchor them to cell membranes. The DHHC domain was discovered in 1999 and named after a conserved sequence motif found in its protein sequence. Roth and colleagues showed that the yeast Akr1p protein could palmitoylate Yck2p in vitro and inferred that the DHHC domain defined a large family of palmitoyltransferases. In mammals twenty three members of this family have been identified and their substrate specificities investigated. Some members of the family such as ZDHHC3 and ZDHHC7 enhance palmitoylation of proteins such as PSD-95, SNAP-25, GAP43, Galphas. Others such as ZDHHC9 showed specificity only toward the H-Ras protein