膜杰作
Star Staining
背景（Background）
Immune checkpoint pathway is a focal point of today’s cancer research. CD47 is one of the best characterized checkpoint proteins. CD47 binding to SIRP alpha aids in tumor evasion of the immune system. Therefore, the pharmaceutical inhibition of CD47 has been considered a promising strategy by many oncologists.
应用说明（Application）
This pair is useful for screening for inhibitors of human CD47 binding to human SIRP alpha.
It is for research use only.
重构方法（Reconstitution）
See Certificate of Analysis for details of reconstitution instruction and specific concentration.
存储（Storage）
Upon receipt, please store all items at -70°C. After reconstitution, the stock solution should be kept at -70°C.
原理（Assay Principles）
This inhibitor screening ELISA pair is designed to facilitate the identification and characterization of new CD47 pathway inhibitors. This assay employs a simple colorimetric ELISA platform, which measures the binding between immobilized human CD47 and in-house developed biotinylated SIRP alpha protein. This product is uniquely suitable for rapid high-throughput screening of putative CD47 inhibitors. Briefly, we provide you with a biotinylated human SIRP alpha protein, a human CD47 protein, an anti-CD47 neutralizing antibody (as method verified Reference), and Streptavidin-HRP reagent.
Your experiment will include 4 simple steps:
1. Coat the plate with human CD47.
2. Mix biotinylated human SIRP alpha and your molecule of interest.
3. Add to the coated human CD47.
4. Add Streptavidin-HRP followed by TMB or other colorimetric HRP substrate.
Finally, the ability of your compound to inhibit CD47: SIRP alpha binding will be determined by comparing OD readings among different experimental groups.
典型数据-Typical Data Please refer to DS document for the assay protocol.
INHIBITION OF CD47: SIRP ALPHA [BIOTINYLATED] BINDING BY ANTI-CD47 NEUTRALIZING ANTIBODY
Serial dilutions of anti-CD47 neutralizing antibody (Catalog # CD7-NA002) (1:2 serial dilutions, from 10 μg/mL to 0.04 μg/mL or 66.67 nM to 0.26 nM) was added into CD47: biotinylated SIRP alpha binding reactions. Background was subtracted from data points prior to log transformation and curve fitting (QC tested).
