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PD-1: PD-L1 [Biotinylated] Inhibitor Screening ELISA Kit

For research use only.

IDComponentsSize
EP101-C01Pre-coated Human PD-1 Microplate1 plate
EP101-C02Anti-PD-1 Neutralizing Antibody100 μL
EP101-C03Human PD-L1-Biotin800 μL
EP101-C04Streptavidin-HRP10 μL
EP101-C0510xWashing Buffer 50 mL
EP101-C06Dilution Buffer50 mL
EP101-C07Substrate Solution12 mL
EP101-C08Stop Solution7 mL

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

Storage/存储

The unopened kit should be stored at 2°C to 8°C. The shelf life is 6 months from the date of receipt. The opened kit should be stored at 2°C to 8°C. The shelf life is 1 month from the date of opening.

Assay Principles/原理

This inhibitor screening ELISA kit 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-L1 to immobilized human PD-1 in a functional ELISA assay, and employs a simple colorimetric sandwich ELISA platform. Briefly, we provide you with a human PD-L1-Biotin protein, a Pre-coated Human PD-1 Microplate, an anti-PD-1 neutralizing antibody (as method verified Std.), and streptavidin-HRP reagent.

Your experiment will include 5 simple steps:

a) All reagents were returned to room temperature (20℃-25℃) before use.

b) Make series the tested sample and control with ditlution buffer, Human PD-L1-Biotin diluted with dilution buffer.

c) Add the diluted sample, Control and the Human PD-L1-Biotin add the plate respectively.

d) Add Streptavidin-HRP followed by TMB or other colorimetric HRP substrate. e) Stop the substrate reaction by add diluted acid. Absorbance (OD) is calculated as the absorbance at 450 nm minus the absorbance at 650 nm to remove background prior to statistical analysis. 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 ELISA

Inhibition of PD-1-PD-L1 [Biotinylated] binding by Anti-PD-1 Neutralizing Antibody measured by competitive ELISA Assay.
Serial dilutions of Anti-PD-1 Neutralizing Antibody (Cat. No. EP101-C02) (1:2 serial dilutions, from 10 μg/mL to 0.02 μg/mL) was added into PD-1 : PD-L1-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. A representative result showed that the IC50 was calculated to be 0.33 μg/mL (QC tested).

Protocol

前沿进展

 
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