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ELISA Kit for B-Cell Activating Factor (BAFF)

CD257; TNFSF13B; BLYS; TALL1; THANK; TNFSF20; ZTNF4; B lymphocyte stimulator; Tumor Necrosis Factor Ligand Superfamily Member 13B; Dendritic cell-derived TNF-like

Specificity

This assay has high sensitivity and excellent specificity for detection of B-Cell Activating Factor (BAFF).
No significant cross-reactivity or interference between B-Cell Activating Factor (BAFF) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant B-Cell Activating Factor (BAFF) and the recovery rates were calculated by comparing the measured value to the expected amount of B-Cell Activating Factor (BAFF) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 98-105 102
EDTA plasma(n=5) 87-102 96
heparin plasma(n=5) 83-90 87

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level B-Cell Activating Factor (BAFF) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level B-Cell Activating Factor (BAFF) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of B-Cell Activating Factor (BAFF) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 82-103% 92-99% 79-102% 87-103%
EDTA plasma(n=5) 79-94% 78-95% 86-96% 94-103%
heparin plasma(n=5) 79-88% 85-101% 98-105% 83-91%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

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Magazine Citations
International Immunopharmacology Expression and effects of B-lymphocyte stimulator and its receptors in T cell-mediated autoimmune arthritis Pubmed:25598291
Int Immunopharmacol BAFF and its receptors involved in the inflammation progress in adjuvant induced arthritis rats PubMed: 26688104
Clinical Rheumatology Metabolic syndrome in Sj PubMed: 26370103
Transplantation Calcineurin Inhibitor Minimization With Ixazomib, an Investigational Proteasome Inhibitor, for the Prevention of Antibody Mediated Rejection in a Preclinical Model. PubMed: 25919767
Rheumatology  Cortical bone density and thickness alterations by high-resolution peripheral quantitative computed tomography: association with vertebral fractures in primary … rheumatology.kew332.abstract
Clinical Immunology Toll-like receptor 9 ligands increase type I interferon induced B-cell activating factor expression in chronic rhinosinusitis with nasal polyposis Pubmed:30056130
Gene A Significant Decrease of BAFF, APRIL, and BAFF Receptors Following Mesenchymal Stem Cell Transplantation in Patients with Refractory Rheumatoid Arthritis Pubmed: 31935514
american journal of physiology-renal physiology The expression of XBP1s in B lymphocytes is critical for pristane-induced lupusnephritis in mice Pubmed: 32249615
diagnostics Interstitial Score and Concentrations of IL-4R¦Á, PAR-2, and MMP-7 in Bronchoalveolar Lavage Fluid Could Be Useful Markers for Distinguishing Idiopathic Interstitial?¡­ 33924683
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