|EP118-C01||High-bind Plate||1 plate|
|EP118-C02||Human BAFF||40 μg|
|EP118-C03||Biotinylated Human BCMA||10 μg|
|EP118-C04||Anti-BCMA Neutralizing Antibody||40 μg|
|EP118-C06||Coating Buffer||12 mL|
|EP118-C07||10xWashing Buffer||50 mL|
|EP118-C08||Blocking Buffer||50 mL|
|EP118-C09-A||ELISA Femto Substrate A||6 mL|
|EP118-C09-B||ELISA Femto Substrate B||6 mL|
TNFRSF17 is a member of the TNF-receptor family, known as tumor necrosis factor receptor superfamily member 17, also known as B cell maturation antigen (BCMA or BCM) or CD269. It is a protein encoded by TNFRSF17 gene. The receptor is expressed on the surface of B lymphocytes and plasma cells and is a marker protein of B lymphocyte maturation.The main ligands of BCMA are BAFF and APRIL. BCMA combined with BAFF could activate the classical and non-classical NF-κB and JNK signaling pathways, up-regulate the expression of anti-apoptotic protein and down-regulate the expression of pro-apoptotic protein. When BCMA binds to APRIL, the expression level of immune checkpoint is up-regulated, creating an immunosuppressed bone marrow microenvironment.
This kit is developed for screening for inhibitors of human BCMA binding to human BAFF.
It is for research use only.
Please see Certificate of Analysis for details of reconstitution instruction and specific concentration.
The unopened kit is stable for 12 months from the date of manufacture if stored at 2°C to 8°C. The opened kit should be stored per components table. The shelf life is 30 days from the date of opening.
This inhibitor screening Assay kit is designed to facilitate the identification and characterization of new BCMA pathway inhibitors. The assay takes advantage of our in house-developed binding of biotinylated human BCMA to immobilized human BAFF in a functional chemiluminescence assay. Briefly, we provide you with a human Biotinylated BCMA protein, a human BAFF protein, an anti-BCMA neutralizing antibody (as method verified Std.), and Streptavidin-HRP reagent.
Your experiment will include 4 simple steps:
a) Coat the plate with human BAFF.
b) Add your molecule of interest to the tests.
c) Add human BCMA-Biotin to bind the coated human BAFF.
d) Add Streptavidin-HRP followed by luminol .
Finally, the half maximal inhibitory concentration (IC50) of your compound to BCMA: BAFF binding will be determined by comparing RLU readings among different experimental groups.
BCMA [BIOTINYLATED]: BAFF BINDING IN THE ABSENCE OF INHIBITORS
Immobilized human BAFF protein at 2 μg/mL (100 μL/well) can bind biotinylated human BCMA with a linear range of 0.0001-0.05 μg/mL when detected by Streptavidin-HRP. Background was subtracted from data points before curve fitting (QC tested).
INHIBITION OF BCMA [BIOTINYLATED]: BAFF BINDING BY ANTI-BCMA NEUTRALIZING ANTIBODY
Serial dilutions of Anti-BCMA Neutralizing antibody (Catalog # EP118-C04) (1:1 serial dilution, from 20 μg/mL to 0.02μg/mL) was added into Biotinylated BCMA: BAFF binding reactions. The assay was performed according to the protocol described below. Background was subtracted from data points prior to log transformation and curve fitting (QC tested).