General 17-Hydroxyprogesterone ,17OHP ELISA Kit

The ELISA is based on the competitive binding enzyme immunoassay technique. The microtiter plate provided in this kit has been pre-coated with an antibody specific to General 17OHP, During the reaction, General 17OHP in the sample or standard competes with a fixed amount of biotin-labeled General 17OHP for sites on a pre-coated Monoclonal antibody specific to General 17OHP. Excess conjugate and unbound sample or standard are washed from the plate. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of General 17OHP in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Precision

Intra-assay Precision (Precision within an assay):Three samples of known concentration were tested twenty times
on one plate to assess intra-assay precision.

Intra-Assay CV: ≤5.7%

Inter-assay Precision (Precision between assays):Three samples of known concentration were tested in five
separate assays to assess inter-assay precision.

Inter-Assay CV: ≤7.9%

RecoveryRecovery was determined by spiking various levels of 17-Hydroxyprogesterone into serum and plasma.

LinearityThe linearity of the kit was assayed by testing samples spiked with appropriate concentration of
17-Hydroxyprogesterone and their serial dilutions. The results were demonstrated by the percentage
of calculated concentration to the expected.