ID | Components | Size |
EP130-C01 | High-bind Plate | 1 plate |
EP130-C02 | Human PD-1 | 10 μg |
EP130-C03 | Biotinylated Human PD-L1 | 20 μg |
EP130-C04 | Anti-PD-1 Neutralizing Antibody | 20 μg |
EP130-C05 | Streptavidin-HRP | 10 μg |
EP130-C06 | Coating Buffer | 12 mL |
EP130-C07 | 10xWashing Buffer | 50 mL |
EP130-C08 | Blocking Buffer | 50 mL |
EP130-C09 | Substrate Solution | 24 mL |
EP130-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)
The inhibitor screening ELISA kit is designed to facilitate the identification and characterization of new PD-1 pathway inhibitors.
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)
Unopened kit should be stored at 2°C -8°C upon receiving. Find the expiration date on the outside packaging and do not use reagents past their expiration date.
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-L1 to immobilized human PD-1 in a functional ELISA assay, and employs a simple colorimetric ELISA platform. Briefly, we provide you with a human Biotinylated PD-L1 protein, a human PD-1 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-1.
b) Add your molecule of interest to the tests.
c) Add human PD-L1-Biotin to bind the coated human PD-1.
d) Add Streptavidin-HRP followed by TMB or other colorimetric HRP substrate.
Finally, the half maximal inhibitory concentration (IC50) of your compound to PD-1: PD-L1 binding will be determined by comparing OD readings among different experimental groups.
典型数据-Typical Data Please refer to DS document for the assay protocol.
INHIBITION OF PD-1: PD-L1 [BIOTINYLATED] BINDING BY ANTI-PD-1 NEUTRALIZING ANTIBODY
Serial dilutions of Anti-PD-1 Neutralizing antibody (Catalog # EP130-C04) (1:1 serial dilution, from 5μg/mL to 0.02μg/mL) was added into PD-1: Biotinylated PD-L1 binding reactions. The assay was performed according to the protocol described below. Background was subtracted from data points prior to log transformation and curve fitting (QC tested).