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ELK Biotechnology

ELK6268

Rat BAFF(B-Cell Activating Factor) ELISA Kit

Rat BAFF(B-Cell Activating Factor) ELISA Kit

Size
Sensitivity
Detection range

Rat BAFF(B-Cell Activating Factor) ELISA Kit

Overview

Product name: Rat BAFF/CD257(B-Cell Activating Factor) ELISA Kit
Reactivity: Rat
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: 27 pg/mL
Standard: 5000 pg/mL
Detection Range: 78.13-5000 pg/mL
Sample Type: Serum, plasma, tissue homogenates, cell lysates and other biological fluids
Assay Length: 3.5h
Research Area: CD & Adhesion molecule;Tumor immunity;Infection immunity;Immune molecule;
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 Rat BAFF/CD257. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Rat 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 Rat 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 Rat BAFF/CD257 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
5000.00 2.132 2.045
2500.00 1.579 1.492
1250.00 1.067 0.980
625.00 0.875 0.788
312.50 0.516 0.429
156.25 0.323 0.236
78.13 0.235 0.148
0.00 0.087 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) 78-96% 87%
EDTA plasma(n=5) 95-107% 101%
Heparin plasma(n=5) 82-95% 88%

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) 85-103% 82-96% 87-105% 90-99%
EDTA plasma(n=5) 79-93% 86-92% 84-98% 89-102%
Heparin plasma(n=5) 85-97% 93-101% 97-103% 91-99%
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