Description
This GLP-1 Total ELISA kit is used for the non-radioactive quantification of GLP-1 (7-36 and 9-36) in serum, plasma, and cell tissue culture. The GLP-1 sequence is highly conserved between the species, with no sequence variation occurring in mammals. One kit is sufficient to measure 39 unknown samples in duplicate.
This assay is a Sandwich ELISA based, sequentially, on: 1) capture of GLP-1 Total molecules from samples to the wells of a microtiter plate coated by a pre-titered amount of anti- GLP-1 polyclonal antibody, 2) wash away of unbound materials from samples, 3) binding of a biotinylated anti- GLP-1 monoclonal antibody to the captured molecules, 4) wash away of unbound materials from samples, 5) conjugation of horseradish peroxidase to the immobilized biotinylated antibodies, 6) wash away of free enzyme conjugates, and 7) quantification of immobilized antibody-enzyme conjugates by monitoring horseradish peroxidase activities in the presence of the substrate 3,3’,5,5’-tetramethylbenzidine. The enzyme activity is measured spectrophotometrically by the increased absorbency at 450 nm, corrected from the absorbency at 590 nm, after acidification of formed products. Since the increase in absorbency is directly proportional to the amount of captured GLP-1 Total 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 GLP-1.
