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PD-1 [Biotinylated] : PD-L1 Inhibitor Screening ELISA Assay Pair

Catalog Components Size (96 tests) Size (480 tests)
A001-214 Human PD-L1 25 μg 125 μg
A002-214 Human PD-1-Biotin 5 μg 25 μg
A003-214 Streptavidin-HRP 10 μg 25 μg
PD1-NA001 Anti-PD-1 Neutralizing Antibody 20 μg 100 μg

Background/背景

Immune checkpoint pathway is a focal point of today’s cancer research. PD-1 is one of the best characterized checkpoint proteins. The binding between PD-1 and its ligand PD-L1 suppresses T-cell activation and allows cancer cells to escape from body’s immune surveillance. Therefore, the pharmaceutical inhibition of PD-1 or its ligand has been considered a promising strategy by many oncologists.

Application/应用

This pair is useful for screening for inhibitors of human PD-1 binding to human PD-L1.

Reconstitution/重构方法

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

Storage/存储

See Certificate of Analysis for details of storage conditions.

Be sure to store each component at the proper temperature upon arrival.

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 PD-1 pathway inhibitors. The assay takes advantage of our in house-developed binding of biotinylated human PD-1 to immobilized human PD-L1 in a functional ELISA assay, and employs a simple colorimetric sandwich ELISA platform. Briefly, we provide you with a human PD-1-Biotin protein, a human PD-L1 protein, an anti-PD-1 neutralizing antibody (as method verified Std.), and streptavidin-HRP reagent. Your experiment will include 4 simple steps:

  • Coat the plate with human PD-L1.
  • Add Human PD-1-Biotin to bind the coated human PD-L1.
  • 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 PD-1 : PD-L1 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 PD-1 to Immobilized Human PD-L1 in a Functional ELISA Assay

Immobilized human PD-L1 protein at 2 μg/mL (100 μL/well) can bind human PD-1-Biotin with a linear range of 0.038 - 0.6 μg/mL when detected by Streptavidin-HRP. Background was subtracted from data points before curve fitting.

Biotinylated Human FACS

Inhibition of PD-1-PD-L1 binding by Anti-PD-1 Neutralizing Antibody (Cat. No. PD1-NA001) measured using the PD-1 [Biotinylated] : PD-L1 Inhibitor Screening ELISA Assay Pair (Cat. No. EP-101)

Serial dilutions of anti-PD-1 neutralizing antibody (Cat. No. PD1-NA001) (1:2 serial dilutions, from 10 μg/mL to 0.078 μg/mL (66.667 nM to 0.521 nM) was added into PD-L1 : PD-1-Biotin 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-101-96tests / ¥6000

EP-101-480tests / ¥21500

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