17alpha-OHP ELISA kit for the quantitative determination of 17α-Hydroxyprogesterone in human serum using a competitive immunoassay format on a 96-well break-apart microplate.
The Instructions for Use (IFU) provided on this website are for informational purposes only. Always refer to the IFU included in the kit for the most current and accurate version.
Regulatory Status
CAN: IVD • EU: IVD CE • USA: IVD. The Instructions for Use (IFU) provided on this website are for informational purposes only.
The 17α-OHP ELISA is a competitive immunoassay. Competition occurs between 17α-OHP present in calibrators, controls, specimen samples and an enzyme-labelled antigen (HRP conjugate) for a limited number of anti-17α-OHP antibody binding sites on the microplate wells.
After a washing step that removes unbound materials, the TMB substrate (enzyme substrate) is added which reacts with HRP to form a blue-coloured product that is inversely proportional to the amount of 17α-OHP present.
Following an incubation, the enzymatic reaction is terminated by the addition of the stopping solution, converting the colour from blue to yellow. The absorbance is measured on a microplate reader at 450 nm. A set of calibrators is used to plot a calibrator curve from which the amount of 17α-OHP in specimen samples and controls can be directly read.
Add 25 µL sample and HRP conjugate to antibody-coated wells
Competitive binding incubation
Wash unbound material, add TMB substrate
Add stop solution, read at 450 nm
Allow all reagents and specimens to reach room temperature. Prepare working wash solution according to instructions.
Dispense 25 µL of calibrators, controls, and specimen samples into the respective antibody-coated microplate wells.
Add the enzyme-labelled antigen (HRP conjugate) to each well. The 17α-OHP in samples competes with the conjugate for limited antibody binding sites.
Cover and incubate to allow competitive binding to occur. The amount of bound conjugate is inversely proportional to the 17α-OHP concentration.
Remove unbound materials by washing the wells with wash solution.
Add TMB substrate to each well. The enzyme reacts with TMB to form a blue-coloured product inversely proportional to 17α-OHP present in the sample.
Add stop solution to terminate the reaction, converting the colour from blue to yellow. Read absorbance at 450 nm on a microplate reader.
Plot a calibrator curve using the calibrator values and directly read the 17α-OHP concentration of specimen samples and controls from the curve.
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