Detection Method:Sandwich
Test principle:This assay employs a two-site sandwich ELISA to quantitate KRAS in samples. An antibody specific for KRAS has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyKRAS present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for KRAS 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 KRAS bound in the initial step. The color development is stopped and the intensity of the color is measured.
Product Overview:Like other members of the Ras family, the KRAS protein is a GTPase and is an early player in many signal transduction pathways. KRAS is usually tethered to cell membranes because of the presence of an isoprenyl group on its C-terminus. When mutated, KRAS is a oncogene. The protein product of the normal KRAS gene performs an essential function in normal tissue signaling, and the mutation of a KRAS gene is an essential step in the development of many cancers.KRAS acts as a molecular on/off switch, once it is turned on it recruits and activates proteins necessary for the propagation of growth factor and other receptors' signal, such as c-Raf and PI 3-kinase. KRAS binds to GTP in the active state and possesses an intrinsic enzymatic activity which cleaves the terminal phosphate of the nucleotide converting it to GDP.
