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Human HLA-B27(Leukocyte Antigen B27) Microsample ELISA Kit
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ELK0864MS
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48 T $320.00
96 T $458.00
96 T*5 $1947.00
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Overview

Product name: Human HLA-B27(Leukocyte Antigen B27) Microsample ELISA Kit
Reactivity: Human
Alternative Names: HALB27; Human leukocyte antigen B27
Assay Type: Sandwich
Sensitivity: 0.17 ng/mL
Standard: 50 ng/mL
Detection Range: 0.78-50 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
50.00 2.254 2.165
25.00 1.634 1.545
12.50 1.167 1.078
6.25 0.806 0.717
3.13 0.549 0.460
1.56 0.369 0.280
0.78 0.235 0.146
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) 93-113% 103%
EDTA plasma(n=5) 94-121% 108%
Heparin plasma(n=5) 90-112% 101%

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) 81-112% 82-120% 91-107% 83-99%
EDTA plasma(n=5) 86-112% 91-99% 83-105% 80-113%
Heparin plasma(n=5) 81-113% 83-96% 81-102% 85-104%

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