Description
This Enzyme Immunoassay kit is designed to detect a specific peptide and its related peptides based on the principle of ÒcompetitiveÓ enzyme immunoassay.
The immunoplate in this kit is pre-coated withsecondary antibody and the nonspecific binding sites are blocked. The secondaryantibody can bind to the Fc fragment of the primary antibody (peptide antibody)whose Fab fragment will be competitively bound by both biotinylated peptide andpeptide standard or targeted peptide in sample. The biotinylated peptide isable to interact with streptavidin-horseradish peroxidase (SA-HRP) whichcatalyzes the substrate solution composed of 3,3Õ, 5,5Õ-tetramethylbenzidine(TMB) and hydrogen peroxidase to produce a blue colored solution. Theenzyme-substrate reaction is stopped by hydrogen chloride (HCI) and thesolution turns to yellow. The intensity of the yellow is directly proportionalto the amount of biotinylated peptide-SA-HRP complex but inversely proportionalto the amount of the peptide in standard solutions or samples. This is due tothe competitive binding of the biotinylated peptide and the peptide in standardsolutions or samples to the peptide antibody (primary antibody). A standardcurve of a peptide with known concentration can be established accordingly. Thepeptide with unknown concentrations in samples can be determined byextrapolation to this standard curve.
