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Human BAFF/CD257(B-Cell Activating Factor) Microsample ELISA Kit
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ELK2145MS
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48 T $320.00
96 T $458.00
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Overview

Product name: Human BAFF/CD257(B-Cell Activating Factor) Microsample ELISA Kit
Reactivity: Human
Alternative Names: CD257; TNFSF13B; BLYS; TALL1; THANK; TNFSF20; ZTNF4; B lymphocyte stimulator; Tumor Necrosis Factor Ligand Superfamily Member 13B; Dendritic cell-derived TNF-like; BAFF
Assay Type: Sandwich
Sensitivity: 29 pg/mL
Standard: 5000 pg/mL
Detection Range: 78.13-5000 pg/mL
Sample Type: serum, plasma, tissue homogenates and other biological fluids
Assay Length: 3.5h
Research Area: CD & Adhesion molecule;Tumor immunity;Infection immunity;Immune molecule;
Uniprot ID: Q9Y275
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 BAFF/CD257. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Human BAFF/CD257. 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 BAFF/CD257, 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 BAFF/CD257 in the samples is then determined by comparing the OD of the samples to the standard curve.

Standard curve

Concentration (pg/mL) OD Corrected OD
5000.00 1.970 1.865
2500.00 1.599 1.494
1250.00 1.074 0.969
625.00 0.803 0.698
312.50 0.537 0.432
156.25 0.314 0.209
78.13 0.223 0.118
0.00 0.105 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 BAFF/CD257 and the recovery rates were calculated by comparing the measured value to the expected amount of BAFF/CD257 in samples.
Matrix Recovery range Average
serum(n=5) 93-99% 96%
EDTA plasma(n=5) 86-126% 106%
Heparin plasma(n=5) 90-110% 100%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of BAFF/CD257 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) 81-102% 92-101% 92-99% 89-111%
EDTA plasma(n=5) 91-119% 93-108% 82-116% 86-116%
Heparin plasma(n=5) 94-101% 94-111% 88-101% 85-120%

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