Description(s):
Brain-type Natriuretic Peptide is a hormone that is produced in ventricular myocytes and is secreted when the heart blood volume expands. The precursor to BNP, proBNP, is cleaved to form N-terminal proBNP (NT-proBNP) and the biologically active polypeptide mature BNP. These two peptides are secreted into the blood in response to pressure overload. NT-proBNP can be used as an indicator to screen for heart failure.
Principle
The wells of the plate are coated with capture antibody specific to NT Pro-BNP. In this assay, the test sample initially reacts with the solid phase capture antibody, resulting in NT Pro-BNP being bound to the well. The wells are then washed to remove unbound proteins, and biotin-linked antibodies are added to bind to the immobilized NT Pro-BNP. After washing away the unbound antibodies, Streptavidin-HRP conjugate is added to form a complex with the antibody-bound NT Pro-BNP. After incubation, the wells are washed to remove unbound Streptavidin-HRP conjugate. TMB substrate is then added and forms a blue color when the HRP-linked antibodies are detected. The reaction is then terminated with Stop Solution, which changes the color from blue to yellow. The NT Pro-BNP concentration in each well is directly proportional to its color intensity and can be quantified by measuring its optical density at 450 nm (OD450) in a microplate reader.
