Abcam’s Amphiregulin Mouse ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of mouse Amphiregulin in serum, plasma and cell culture supernatants.
This assay employs an antibody specific for mouse Amphiregulin coated on a 96-well plate. Standards and samples are pipetted into the wells and Amphiregulin present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-mouse Amphiregulin antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of Amphiregulin bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.
The mature secreted form of amphiregulin is an 84-amino acid residue glycosylated polypeptide which is generated by proteolytic processing of a 252-amino acid transmembrane precursor. Seven different polypeptide ligands, which derive from distinct genes, are capable of binding to the extracellular domain of EGFR; these are EGF, TGFa, AR, HB-EGF, cripto-1, epiregulin, and betacellulin. All of these growth factors contain a characteristic EGF-like domain which is defined by 6 evenly spaced cysteine residues that generate 3 loops through the formation of disulfide bonds.
Amphiregulin expression is induced by phorbol ester, estrogen, androgen, and other EGFR ligands. In vitro, amphiregulin functions as an autocrine growth stimulator to drive proliferation of colon carcinoma cells, normal and oncogene-transformed mammary epithelial cells, cervical carcinoma cells, prostate cancer cells, and keratinocytes. Amphiregulin is important in the development of human placenta and murine mammary gland. It is reportedly overexpressed in human cancers of breast, colon, stomach and pancreas.