Detection Method:Sandwich
Test principle:This assay employs a two-site sandwich ELISA to quantitate PIM3 in samples. An antibody specific for PIM3 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyPIM3 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for PIM3 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 PIM3 bound in the initial step. The color development is stopped and the intensity of the color is measured.
Product Overview:The deduced 326-amino acid protein has a calculated molecular mass of 35.9 kD. PIM3 shares 95% amino acid identity with rodent Pim3 and 57.1% and 44.0% amino acid identity with human PIM1 and PIM2 , respectively.
Northern blot analysis detected a 2.4-kb transcript in heart, skeletal muscle, brain, spleen, kidney, placenta, lung, and peripheral blood leukocytes. Expression was not detected in colon, thymus, liver, and small intestine. Immunohistochemical analysis showed no expression of PIM3 in normal liver, but it localized diffusely in large regenerative nodules and adenomatous hyperplasia, lesions with precancerous potential, adjacent to hepatocellar carcinoma areas. PIM3 was expressed in all hepatoma cell lines examined.
