Detection Method:Sandwich
Test principle:This assay employs a two-site sandwich ELISA to quantitate PDIA6 in samples. An antibody specific for PDIA6 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyPDIA6 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for PDIA6 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 PDIA6 bound in the initial step. The color development is stopped and the intensity of the color is measured.
Product Overview:Protein disulfide isomerases, such as PDIA6, are endoplasmic reticulum resident proteins that catalyze formation, reduction, and isomerization of disulfide bonds in proteins and are thought to play a role in folding of disulfide-bonded proteins The deduced 440-amino acid protein has a calculated molecular mass of 48.1 kD. It has a putative N-terminal signal sequence, followed by 2 thioredoxin like domains and a C-terminal ER retention signal. Mutation analysis revealed that the first thioredoxin-like motif of P5 was more important than the second for isomerase activity, and that the first cysteine in each motif was necessary for isomerase activity. Thioredoxin motif mutants of P5 lacking isomerase activity retained chaperone activity with citrate synthase as substrate, indicating that, like PDI, the isomerase and chaperone activities of P5 are likely independent.
