Detection Method:Sandwich
Test principle:This assay employs a two-site sandwich ELISA to quantitate DIRAS3 in samples. An antibody specific for DIRAS3 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyDIRAS3 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for DIRAS3 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 DIRAS3 bound in the initial step. The color development is stopped and the intensity of the color is measured.
Product Overview:ARHI contains a highly conserved GTP-binding domain, a putative effector domain distinct from that of RAS and RAP proteins, and a C-terminal membrane localization motif. Northern blot analysis detected a 1.9-kb ARHI transcript in all normal breast and ovarian epithelial cell cultures tested, as well as in normal ovary, heart, liver, pancreas, and brain. Expression was absent in nearly all breast and ovarian cancer cell lines and all primary ovarian cancer cell lines tested. Western blot analysis detected a 26-kD ARHI protein in all normal breast and ovarian cell lines but not in any breast and ovarian cancer cell lines tested. Expression of ARHI in breast and ovarian cancer cell lines but not in lung cancer cell lines led to growth inhibition. Stimulation of normal cell lines with growth factors led to decreased expression of ARHI as well as the cell growth inhibition-associated protein WAF1.
