Detection Method:Sandwich
Test principle:This assay employs a two-site sandwich ELISA to quantitate PLK1 in samples. An antibody specific for PLK1 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyPLK1 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for PLK1 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 PLK1 bound in the initial step. The color development is stopped and the intensity of the color is measured.
Product Overview:Plk1 consists of 603 amino acids and is 66kDa. In addition to the N-terminus kinase domain, there are two conserved polo-box regions of 30 amino acids at the C-terminus.
Plk1 is an early trigger for G2/M transition. Plk1 supports the functional maturation of the centrosome in late G2/early prophase and establishment of the bipolar spindle. Plk1 phosphorylates and activates cdc25C, a phosphatase that dephosphorylates and activates the cyclinB/cdc2 complex. Plk phosphorylates and activates components of the APC. The APC, which is activated by Fizzy-Cdc20 family proteins, is a cell cycle ubiquitin-protein ligase (E3) that degrades mitotic cyclins, chromosomal proteins that maintain cohesion of sister chromatids, and anaphase inhibitors.
