Detection Method:Sandwich
Test principle:This assay employs a two-site sandwich ELISA to quantitate Ub in samples. An antibody specific for Ub has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyUb present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for Ub 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 Ub bound in the initial step. The color development is stopped and the intensity of the color is measured.
Product Overview:Posttranslational modification of proteins by the addition of the small protein SUMO, or sumoylation, regulates protein structure and intracellular localization. SAE1 and UBA2 form a heterodimer that functions as a SUMO-activating enzyme for the sumoylation of proteins.
The deduced SAE1 and SAE2 proteins contain 347 and 640 amino acids, respectively. SAE1 shares sequence similarity with the N terminus of ubiquitin-activating E1 enzymes, and SAE2 share sequence similarity with the C terminus of E1 enzymes. Both SAE subunits contain a conserved nucleotide-binding motif, and SAE2 contains an E1-like active-site cysteine. SAE2 has a calculated molecular mass of 72 kD. It had an apparent molecular mass of 90 kD by SDS-PAGE.
