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| ID | Components | Size |
| 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-C05 | Streptavidin-HRP | 10 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 |
Background/背景
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.
Application/应用
This kit is developed for screening for inhibitors of human BCMA binding to human BAFF.
It is for research use only.
Reconstitution/重构方法
Please see Certificate of Analysis for details of reconstitution instruction and specific concentration.
Storage/存储
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.
Assay Principles/原理
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.
Typical Data/典型数据
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).
Protocol
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).
Protocol
