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Human HLA-B27(Leukocyte Antigen B27) ELISA Kit
ELK0864
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48 T $320.00
96 T $458.00
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Overview

Product name: Human HLA-B27(Leukocyte Antigen B27) ELISA Kit
Reactivity: Human
Alternative Names: HALB27; Human leukocyte antigen B27
Assay Type: Sandwich
Sensitivity: 0.12 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: Immune molecule
Uniprot ID: O19193
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 HLA-B27. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Human HLA-B27. 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 HLA-B27, 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 HLA-B27 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 2.195 2.106
10.00 1.657 1.568
5.00 1.136 1.047
2.50 0.806 0.717
1.25 0.539 0.450
0.63 0.364 0.275
0.32 0.236 0.147
0.00 0.089 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 HLA-B27 and the recovery rates were calculated by comparing the measured value to the expected amount of HLA-B27 in samples.
Matrix Recovery range Average
serum(n=5) 87-95% 90%
EDTA plasma(n=5) 96-107% 102%
Heparin plasma(n=5) 81-97% 89%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of HLA-B27 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) 97-105% 88-95% 96-102% 83-97%
EDTA plasma(n=5) 89-103% 85-93% 89-97% 95-102%
Heparin plasma(n=5) 85-98% 81-96% 87-98% 92-103%