Catalog No: RE10139
17-OHP(17-Hydroxyprogesterone) ELISA Kit
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48T

$299.00

96T

$399.00

Product Details

Species:

Universal

Alternative Name:

17OHP; 17-OH Progesterone; 17-Alpha-Hydroxyprogesterone

Assay Type:

Competitive

unit:

pg/mL

Standard:

20000 pg/mL

Sensitivity:

87.3 pg/mL

Detection Range:

312.5-20000 pg/mL

Assay Time:

1.5H

Sample Type:

Serum, Plasma, Tissue homogenates, Cell lysates, Cell culture supernates and Other biological fluids

Research:

Endocrinology;Reproductive science;Hormone metabolism;

Test Principle:

This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with 17-OHP. Standards or samples are added to the appropriate micro ELISA 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.

Technical Data

As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.
pg/mL OD Corrected
20000 0.196
10000 0.332
5000 0.547
2500 0.881
1250 1.309
625 1.701
312.5 2.077
0 2.517

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level were tested on 3 different plates, 20 replicates in each plate, respectively.

Intra-assay Precision Inter-assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean(ng/mL) 393.7 840.34 5617.98 409.84 628.93 6993.01
Standard deviation 41.95 92.71 290.18 52.22 115.4 361.21
C V (%) 5.09 5.4 4.56 4.55 4.74 2.43

Rate of recovery

The recovery of spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
Sample Type Range(%) Average Recovery(%)
Serum (n=8) 80-92 86
EDTA plasma (n=8) 90-105 97
Cell culture media (n=8) 88-102 95

Linear

Samples were spiked with high concentrations of target proteins and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.
Serum (n=5) EDTA plasma (n=5) Cell culture media (n=5)
1:2 Range (%) 87-95 80-91 86-99
Average (%) 89 80 86
1:4 Range (%) 85-96 80-91 92-104
Average (%) 87 84 92
1:8 Range (%) 92-102 89-103 82-96
Average (%) 95 91 92
1:16 Range (%) 81-92 82-95 95-102
Average (%) 90 89 100

Assay Procedures

Description