The DiaSource 17OH-Progesterone ELISA kit is a solid phase Enzyme Linked Immunosorbent Assay performed on microtiterplates.
During a 1 hour incubation step, at room temperature, with shaking, 17-OHP present in calibrators, controls and samples competes with 17-OHP-HRP conjugate for binding sites of a specific antibody immobilized on the wells of the microtiterplate. After this incubation, microtiterplate is washed to stop the competition reaction.
The Chromogenic solution (TMB) is added and incubated for 30 minutes. 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 inversely proportional to the 17-OHP concentration.
A calibration curve is plotted and the 17-OHP concentrations of the samples are determined by dose interpolation from the calibration curve.