Detection Method:Sandwich
Test principle:This assay employs a two-site sandwich ELISA to quantitate PPCDC in samples. An antibody specific for PPCDC has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyPPCDC present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for PPCDC 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 PPCDC bound in the initial step. The color development is stopped and the intensity of the color is measured.
Product Overview:Biosynthesis of coenzyme A (CoA) from pantothenic acid (vitamin B5) is an essential universal pathway in prokaryotes and eukaryotes. PPCDC (EC 4.1.1.36), one of the last enzymes in this pathway, converts phosphopantothenoylcysteine to 4-prime-phosphopantetheine. PPCDC functioned within the CoA synthetic pathway, and they verified PPCDC function by complementation in E. coli. Incubation of PPCS, PPCDC, and the bifunctional enzyme COASY with the necessary substrates and cofactors reconstituted the 4-step biochemical transformation of phosphopantothenate to CoA. Saccharomyces cerevisiae?Hal3 and Vhs3 are moonlighting proteins, forming an atypical heterotrimeric decarboxylase (PPCDC) required for CoA biosynthesis, and regulating cation homeostasis by inhibition of the Ppz1 phosphatase.
