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Human anti-21-OH Ab(anti 21-hydroxylase antibody) ELISA Kit
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ELK10071
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48 T $320.00
96 T $458.00
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Overview

Product name: Human anti-21-OH Ab(anti 21-hydroxylase antibody) ELISA Kit
Reactivity: Human
Alternative Names: 21-OH Ab
Assay Type: Sandwich
Sensitivity: 0.189 ng/mL
Standard: 20 ng/mL
Detection Range: 0.32-20 ng/mL
Sample type: serum, plasma,Tissue homogenates and other biological fluids.
Assay length: 3.5h
Research Area: Immunology
Test principle: The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Human 21-OH Ab. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Human 21-OH Ab. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Human 21-OH Ab, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. 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 Human 21-OH Ab 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
20.00 2.258 2.165
10.00 1.587 1.494
5.00 1.136 1.043
2.50 0.757 0.664
1.25 0.496 0.403
0.63 0.326 0.233
0.32 0.152 0.059
0.00 0.093 0.000

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 21-OH Ab and the recovery rates were calculated by comparing the measured value to the expected amount of 21-OH Ab in samples.
Matrix Recovery range Average
serum(n=5) 96-108% 102%
EDTA plasma(n=5) 87-102% 94%
Heparin plasma(n=5) 82-95% 88%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of 21-OH Ab 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) 85-97% 87-96% 89-103% 82-95%
EDTA plasma(n=5) 81-97% 82-96% 95-102% 85-92%
Heparin plasma(n=5) 85-94% 87-101% 87-101% 82-93%