Detection Method:Sandwich
Test principle:This assay employs a two-site sandwich ELISA to quantitate WWTR1 in samples. An antibody specific for WWTR1 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyWWTR1 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for WWTR1 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 WWTR1 bound in the initial step. The color development is stopped and the intensity of the color is measured.
Product Overview:Using yeast 2-hybrid analysis of a HeLa cDNA library with XPA as bait, followed by RACE PCR, Nakatsu et al. (2000) cloned XAB2. The 855-amino acid protein has 3 acidic regions, 15 class I tetratricopeptide repeats, and a conserved C-terminal region.By in vitro pull-down assay, Nakatsu et al. (2000) confirmed the specific interaction of XPA and XAB2 in yeast. Coimmunoprecipitation assays showed that XAB2 also interacts with CSA (ERCC8), CSB (ERCC6) and RNA polymerase II both in vitro and in vivo. Microinjection of XAB2 antibodies in primary fibroblasts and fibroblasts from XPC patients inhibited mRNA synthesis and transcription-coupled repair (TCR), but did not affect global genome repair (GGR), suggesting that XAB2 functions in normal transcription and in TCR.
