Description
This kit is for non-radioactive quantification of biologically active forms of Glucagon-Like Peptide-1 [i.e. GLP-1 (7-36 amide) and GLP-1 (7-37)] in plasma and other biological media. It is highly specific for the immunologic measurement of active GLP-1 and will not detect other forms of GLP-1 (e.g., 1-36 amide, 1-37, 9-36 amide, or 9-37). The GLP-1 sequence is highly conserved between the species, with no sequence variation occurring at all in mammals. One kit is sufficient to measure 39 unknown samples in duplicate.
This assay is based, sequentially, on: 1) capture of active GLP-1 from sample by a monoclonal antibody, immobilized in the wells of a microwell plate, that binds specifically to the N-terminal region of active GLP_1 molecule, 2) washing to remove unbound materials, 3) binding of an anti GLP-1-alkaline phosphatase detection conjugate to the immobilized GLP-1, 4) washing off unbound conjugate, and 5 ) quantification of bound detection conjugate by adding MUP (methyl umbelliferyl phosphate) which in the presence of alkaline phosphatase forms the fluorescent product umbelliferone. Since the amount of fluorescence generated is directly proportional to the concentration of active GLP-1 in the unknown sample, the latter can be derived by interpolation from a reference curve generated in the same assay with reference standards of known concentrations of active GLP-1.
