Description
Immunoenzymetric assay for the in vitro quantitative measurement of the human Insulin-like Growth Factor Binding Protein-3 (IGFBP-3) in serum.
The IGFBP3-ELISA is a solid phase Enzyme Amplified Sensitivity Immunoassay performed on microtiterplates. Standards and samples react with the capture monoclonal antibody (MAb 1) coated on microtiter well and with a monoclonal antibody (MAb 2) labelled with horseradish peroxidase (HRP). After an incubation period allowing the formation of a sandwich: coated MAb 1 – human IGFBP3Ð MAb 2 – HRP, the microtiterplate is washed to remove unbound enzyme labelled antibody.Bound enzyme-labelled antibody is measured through a chromogenic reaction. Chromogenic solution (TMB) is added and incubated. The reaction is stopped with the addition of Stop Solution and the microtiterplate is then read at the appropriate wavelength. The amount of substrate turnover is determined colourimetrically by measuring the absorbance, which is proportional to the human IGFBP3 concentration.A calibration curve is plotted and IGFBP3 concentration in samples is determined by interpolation from the calibration curve.
