ID | Components | Size |
EP101-C01 | High-bind Plate | 1 plate |
EP101-C02 | Human PD-L1 | 25 μg |
EP101-C03 | Anti-PD-1 Neutralizing Antibody | 20 μg |
EP101-C04 | Human PD-1-Biotin | 5 μg |
EP101-C05 | Streptavidin-HRP | 10 μg |
EP101-C06 | Coating Buffer | 12 mL |
EP101-C07 | 10xWashing Buffer | 50 mL |
EP101-C08 | Blocking Buffer | 50 mL |
EP101-C09 | Substrate Solution | 12 mL |
EP101-C10 | Stop Solution | 7 mL |
背景(Background)
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.
应用说明(Application)
This kit is developed for screening for inhibitors of human PD-1 binding to human PD-L1.
It is for research use only.
重构方法(Reconstitution)
Please see Certificate of Analysis for details of reconstitution instruction and specific concentration.
存储(Storage)
The unopened kit is stable for 12 months from the date of manufacture if stored at 2°C to 8°C.
原理(Assay Principles)
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-1 to immobilized human PD-L1 in a functional ELISA assay, and employs a simple colorimetric sandwich ELISA platform.
Your experiment will include 4 simple steps:
a) Coat the plate with human PD-L1.
b) Add your molecule of interest to the tests.
c) Add human PD-1-Biotin to bind the coated human PD-L1.
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.
典型数据-Typical Data
INHIBITION OF PD-1-PD-L1 BINDING BY ANTI-PD-1 NEUTRALIZING ANTIBODY (CATALOG # EP101-C03) MEASURED USING THE PD-1 [BIOTINYLATED] : PD-L1 INHIBITOR SCREENING ELISA ASSAY PAIR (CATALOG # EP-101).
Anti-PD-1 neutralizing antibody was diluted from 10 μg/mL to 0.078 μg/ mL (69.628 nM to 0.544 nM) and loaded onto the plate coated by human PD-L1 in the presence of human PD-1-Biotin. Assay was performed according to the protocol in PD-1[Biotinylated] : PD-L1 Inhibitor Screening ELISA Assay Pair. Background was subtracted from data points prior to log transformation and curve fitting (QC tested).
Protocol