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ELK Biotechnology

ELK7875

General 17-OHP(17-Hydroxyprogesterone) ELISA Kit

General 17-OHP(17-Hydroxyprogesterone) ELISA Kit

Size
Sensitivity
Detection range

General 17-OHP(17-Hydroxyprogesterone) ELISA Kit

Overview

Product name: 17-OHP(17-Hydroxyprogesterone) ELISA Kit
Reactivity: General
Alternative Names: 17OHP; 17-OH Progesterone; 17-Alpha-Hydroxyprogesterone
Assay Type: Competitive Inhibition
Sensitivity: 87.3 pg/mL
Standard: 20000 pg/mL
Range: 312.5-20000 pg/mL
Sample Type: serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
Assay Length: 2h
Research Area: Endocrinology;Reproductive science;Hormone metabolism;
Test principle: This assay employs the competitive inhibition enzyme immunoassay technique. The microtiter plate provided in this kit has been pre-coated with 17-OHP protein. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to 17-OHP. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of 17-OHP in the samples is then determined by comparing the OD of the samples to the standard curve.

Standard curve

Concentration (ng/mL) OD Corrected OD
20000.00 0.215 —
10000.00 0.375 —
5000.00 0.527 —
2500.00 0.899 —
1250.00 1.157 —
625.00 1.537 —
312.50 1.887 —
0.00 2.232 —

Precision

Intra-assay Precision (Precision within an assay)：CV%<8%

Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.

Inter-assay Precision (Precision between assays)：CV%<10%

Three samples of known concentration were tested in forty separate assays to assess inter-assay precision.

Recovery

Matrices listed below were spiked with certain level of recombinant 17-OHP and the recovery rates were calculated by comparing the measured value to the expected amount of 17-OHP in samples.
Matrix Recovery range Average
serum(n=5) 90-105% 97%
EDTA plasma(n=5) 86-99% 92%
Heparin plasma(n=5) 80-95% 86%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of 17-OHP and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Matrix 1:2 1:4 1:8 1:16
serum(n=5) 78-92% 83-96% 91-103% 96-105%
EDTA plasma(n=5) 83-96% 92-104% 89-101% 85-97%
Heparin plasma(n=5) 85-94% 91-103% 85-97% 87-96%
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