BAFFR [Biotinylated] : BAFF Inhibitor Screening ELISA Kit

BAFFR [Biotinylated] : BAFF Inhibitor Screening ELISA Kit

Background/背景

BAFF receptor (B-cell activating factor receptor, BAFF-R), also known as tumor necrosis factor receptor superfamily member 13C (TNFRSF13C), is a membrane protein of the TNF receptor superfamily which recognizes BAFF. B-cell activating factor (BAFF) enhances B-cell survival in vitro and is a regulator of the peripheral B-cell population. Overexpression of BAFF in mice results in mature B-cell hyperplasia and symptoms of systemic lupus erythematosus (SLE). Also, some SLE patients have increased levels of BAFF in serum. Therefore, it has been proposed that abnormally high levels of BAFF may contribute to the pathogenesis of autoimmune diseases by enhancing the survival of autoreactive B cells.

Application/应用

This kit is developed for screening for inhibitors of human BAFFR 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 ELISA kit is designed to facilitate the identification and characterization of new BAFFR pathway inhibitors. The assay takes advantage of our in house-developed binding of biotinylated human BAFFR to immobilized human BAFF in a functional ELISA assay, and employs a simple colorimetric ELISA platform. Briefly, we provide you with a human Biotinlated BAFFR protein, a human BAFF protein, an anti-BAFFR 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 BAFFR-Biotin to bind the coated human BAFF.

d) Add Streptavidin-HRP followed by TMB or other colorimetric HRP substrate.

Finally, the half maximal inhibitory concentration (IC50) of your compound to BAFFR: BAFF binding will be determined by comparing OD readings among different experimental groups.

 

Typical Data/典型数据

BAFFR [BIOTINYLATED]: BAFF BINDING IN THE ABSENCE OF INHIBITORS
Immobilized human BAFF protein at 1 μg/mL (100 μL/well) can bind biotinylated human BAFFR with a linear range of 0.0002-0.025 μg/mL when detected by Streptavidin-HRP. Background was subtracted from data points before curve fitting (QC tested).

Protocol

INHIBITION OF BAFFR [BIOTINYLATED]: BAFF BINDING BY ANTI-BAFFR NEUTRALIZING ANTIBODY
Serial dilutions of Anti-BAFFR Neutralizing antibody (1:1 serial dilution, from 10 μg/mL to 0.02μg/mL) was added into BAFF: Biotinlated BAFFR binding reactions. The assay was performed according to the protocol described above. Background was subtracted from data points prior to log transformation and curve fitting (QC tested).

Proto

 

特别提示：本公司的所有产品仅可用于科研实验，严禁用于临床医疗及其他非科研用途！