|ID||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|
This pair is useful for screening for inhibitors of human PD-1 binding to human PD-L1.
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.
This product is stable 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.
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:
a) Coat the plate with human PD-L1.
b) Add Human PD-1-Biotin to bind the coated human PD-L1.
c) Add your molecule of interest to the tests.
d) 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.
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.
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.
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