The Human c-Met (Total) ELISA Kit is a solid-phase sandwich Enzyme-Linked Immunosorbent Assay (ELISA) designed to detect and quantify the level of c-Met (Total) in fresh or frozen human cell lysates. The assay recognizes both natural and recombinant full length human c-Met (Total), independent of its phosphorylation state.
Principle of the method
A monoclonal capture antibody specific for c-Met (Total) has been coated onto the wells of the 96-well plate. During the first incubation, standards of known content and unknown samples are pipetted into the wells and the antigen binds to the immobilized (capture) antibody. After washing, a rabbit antibody specific for the target protein is added to the wells and serves as a detection antibody by binding to the immobilized protein captured during the first incubation. After washing, a horseradish peroxidase labeled anti-rabbit IgG is added. This binds to the detection antibody to complete the four member sandwich. After a third incubation and washing to remove all the unbound enzyme, a substrate solution (TMB) is added, which is acted upon by the bound enzyme to produce color. The intensity of this colored product is directly proportional to the concentration of target protein present in the original specimen and the optical density can be read on a standard microplate reader.
Each manufactured lot of this ELISA kit is quality tested for criteria such as sensitivity, specificity, precision, and lot-to-lot consistency. See manual for more information on validation.
Hepatocyte growth factor receptor (HGF R), a product of the proto-oncogene c-Met, is a heterodimeric transmembrane glycoprotein that is a receptor-type tyrosine kinase. The c-Met heterodimer is composed of an alpha chain that is disulfide-linked to a beta chain. Each alpha and beta subunit heterodimer contain 1152 amino acid residues with a calculated molecular mass of approximately 129 kDa. The alpha chain is exposed to the cell surface and the beta chain spans the plasma membrane. c-Met is synthesized as a single-chain precursor which undergoes cotranslational glycosylation and proteolytic cleavage producing the heterodimeric mature form. Human and mouse HGF receptors share 89% amino acid identity. HGF is the ligand for the HGF receptor.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.