Detection Method:Sandwich
Test principle:This assay employs a two-site sandwich ELISA to quantitate EGLN1 in samples. An antibody specific for EGLN1 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyEGLN1 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for EGLN1 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 EGLN1 bound in the initial step. The color development is stopped and the intensity of the color is measured.
Product Overview:EGLN1 catalyzes the post-translational formation of 4-hydroxyproline in hypoxia-inducible factor (HIF) alpha proteins. HIF is a transcriptional complex that plays a central role in mammalian oxygen homeostasis. This protein functions as a cellular oxygen sensor, and under normal oxygen concentration, modification by prolyl hydroxylation is a key regulatory event that targets HIF subunits for proteasomal destruction via the von Hippel-Lindau ubiquitylation complex. The genes encoding these proteins were cloned and termed PHD1, PHD2, and PHD3 by the authors. Direct modulation of recombinant enzyme activity by graded hypoxia, iron chelation, and cobaltous ions mirrored the characteristics of HIF induction in vivo, fulfilling requirements for these enzymes being oxygen sensors that regulate HIF.
