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CD47: SIRP alpha [Biotinylated] Inhibitor Screening ELISA Assay Pair

Catalog Components Size (96 tests) Size (480 tests)
A004-214 Human CD47 35 μg 160 μg
A005-214 Biotinylated Human SIRP alpha 10 μg 10 μg
A003-214 Streptavidin-HRP 10 μg 10 μg
CD7-NA002 Anti-CD47 Neutralizing Antibody 20 μg 100 μg

Background/背景

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.

Application/应用

This pair is useful for screening for inhibitors of human CD47 binding to human SIRP alpha.

Reconstitution/重构方法

See Certificate of Analysis for details of reconstitution instruction and specific concentration.

Storage/存储

Upon receipt, please store all items at -70℃.

After reconstitution, the stock solution should be kept at -70℃.

It is recommended not to freeze thaw more than 3 times.

No activity loss is observed after storage at:

  • Room temperature (RT) for 1 month in lyophilized state;
  • -20℃ for 1 year in lyophilized state;
  • -70℃ for 6 months under sterile conditions after reconstitution.

Assay Principles/原理

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:

  • Coat the plate with human CD47.
  • Mix biotinylated human SIRP alpha and your molecule of interest.
  • Add to the coated human CD47.
  • 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.

 

Typical Data/典型数据

Biotinylated Human FACS

Binding of Biotinylated Human SIRP alpha to Immobilized Human CD47 in a Functional ELISA Assay.

Immobilized human CD47 protein at 3 μg/mL (100 μL/well) can bind biotinylated human SIRP alpha with a linear range of 0.004-0.125 μg/mL when detected by Streptavidin-HRP. Background was subtracted from data points before curve fitting.

Biotinylated Human FACS

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) were added into CD47: biotinylated SIRP alpha binding reactions. The assay was performed according to the above described protocol. Background was subtracted from data points prior to log transformation and curve fitting.

 

前沿进展

货号/价格

EP-102-96tests / ¥6000

EP-102-480tests / ¥21500

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CD47靶点信息
英文全称:Cluster of Differentiation 47
中文全称:分化群47
种类:Homo sapiens
上市药物数量:0详情
临床药物数量:12详情
最高研发阶段:临床二期
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