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Multiplex Assay Kit for B-Cell Activating Factor (BAFF) ,etc. by FLIA (Flow Luminescence Immunoassay)

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

(Note: Up to 8-plex in one testing reaction)

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 78-95 90
EDTA plasma(n=5) 84-98 90
heparin plasma(n=5) 96-104 101
sodium citrate plasma(n=5) 88-99 95

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level B-Cell Activating Factor (BAFF) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) 80-99% 81-95% 81-93% 87-101%
EDTA plasma(n=5) 89-104% 89-105% 82-104% 88-102%
heparin plasma(n=5) 84-98% 93-101% 90-98% 79-101%
sodium citrate plasma(n=5) 96-103% 85-101% 80-88% 82-92%

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
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:BAFF) 1 Analysis buffer 1×20mL
Pre-Mixed Detection Reagent A 1×120μL Assay Diluent A 1×12mL
Detection Reagent B (PE-SA) 1×120μL Assay Diluent B 1×12mL
Sheath Fluid 1×10mL Wash Buffer (30 × concentrate) 1×20mL
Instruction manual 1

Assay procedure summary

1. Preparation of standards, reagents and samples before the experiment;
2. Add 100μL standard or sample to each well,
    add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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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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