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Applications: Screen small molecule inhibitors or antibodies that block BAFF binding to BAFFR.
Background: The BAFF (TNF family ligand B-cell Activating Factor), also known as BLyS, TALL-1 or CD257, is encoded by the TNFSF13B gene and is a Type II membrane-bound protein, which can be released as a soluble ligand upon proteolytic processing. This cytokine is a ligand for the receptors Transmembrane Activator and CAML Interactor (TACI or TNFRSF13B), BAFF Receptor (BAFFR, or TNFRSF13C) and B-cell Maturation Antigen (BCMA, TNFRSF17). These interactions promote cell survival and play a crucial role in B cell development. In particular, BAFFR signaling is an important player in the later stages of B cell differentiation, including the survival of long-lived bone marrow plasma cells and likely for the survival of plasma blasts. BAFF signaling is also required for CCL (chronic lymphocytic leukemia) cell survival. Binding of BAFF to BAFFR results in the degradation of TRAF3 (TNF receptor associated factor 3), which stops the inhibition of SYK and results in activation of pathways necessary for B cells. It also activates the non-canonical slow NF-κB2-dependent pathway, which relies on the activation of NIK (NF-κB-inducing kinase). Targeting BAFF-BAFFR will open new therapeutic avenues in diseases related to these proteins.
Contraindications: The DMSO concentration in the final reaction should be ‰¤1%.
Description: The BAFF:BAFFR [Biotinylated] Inhibitor Screening Chemiluminescence Assay Kit is designed for screening and profiling molecules that block the binding between BAFF (B-cell Activating Factor) and BAFFR (BAFF receptor). This kit comes in a convenient 96-well format, with enough recombinant human biotin-labeled BAFFR (amino acids 23-71), BAFF (amino acids 134-285), streptavidin-HRP, and assay buffer for 100 reactions.A 96-well plate is coated with BAFF protein. After blocking, the plate is pre-incubated with an inhibitor or neutralizing antibody. After incubation with Biotin-BAFFR, the plate is washed and Streptavidin-HRP is added. The ELISA ECL substrate is added, and the resulting signal can be measured using a chemiluminescence microplate reader. The chemiluminescence signal is proportional to the binding of BAFF to BAFFR.
Storage Stability: This assay kit will perform optimally for up to 6 months from date of receipt when the materials are stored as directed.
Uniprot: Q9Y275
Warnings: Avoid freeze/thaw cycles
Biosafety Level: Not applicable (BSL-1)
References: Thompson J. S., et al., 2000 J. Exp. Med. 192(1): 129-136.
Bossen C., et al., 2006 Semin. Immunol. 18(5): 263-275.
Scheighoffer E. and Tybulewicz V., 2021 Curr Opin Immunol. 71:124-131.
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