ID | Components | Size |
EP134-C01 | High-bind Plate | 1 plate |
EP134-C02 | Human B7-1 | 25 μg |
EP134-C03 | Biotinylated Human PD-L1 | 15 μg |
EP134-C04 | Anti-PD-L1 Neutralizing Antibody | 20 μg |
EP134-C05 | Streptavidin-HRP | 5 μg |
EP134-C06 | Coating Buffer | 12 mL |
EP134-C07 | 10xWashing Buffer | 50 mL |
EP134-C08 | Blocking Buffer | 50 mL |
EP134-C09 | Substrate Solution | 12 mL |
EP134-C10 | Stop Solution | 7 mL |
背景(Background)
Programmed death molecule 1 and its ligand (PD-1/PD-L1) are a pair of negative immune costimulatory molecules. Under normal circumstances, the combination of PD-L1 on tissue cell surface and PD-1 on lymphocyte surface can inhibit lymphocyte function and induce apoptosis of activated lymphocyte, thus playing an autoimmune tolerance role. However, pD-L1 is also expressed on the surface of various tumor cells. Pd-l1 expressed by tumor cells can bind to PD-1 on the surface of corresponding lymphocytes, inhibit the function of lymphocytes and the release of cytokines, and induce apoptosis of lymphocytes, leading to immune escape of tumor cells. Therefore, blocking the PD-1/PD-L1 pathway can enhance lymphocyte activity to achieve the effect of tumor immunotherapy.
应用说明(Application)
This kit is developed for screening for inhibitors of human PD-L1 binding to human B7-1.
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.
The opened kit should be stored per components table. The shelf life is 30 days from the date of opening.
原理(Assay Principles)
This inhibitor screening ELISA kit is designed to facilitate the identification and characterization of new PD-L1 pathway inhibitors. The assay takes advantage of our in house-developed binding of biotinylated human PD-L1 to immobilized human B7-1 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 B7-1 protein, an anti-PD-L1 neutralizing antibody (as method verified Std.), and Streptavidin-HRP reagent.
Your experiment will include 4 simple steps:
a) Coat the plate with human B7-1.
b) Add your molecule of interest to the tests.
c) Add human PD-L1-Biotin to bind the coated human B7-1.
d) Add Streptavidin-HRP followed by TMB or other colorimetric HRP substrate.
Finally, the half maximal inhibitory concentration (IC50) of your compound to PD-L1: B7-1 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-L1 [Biotinylated]: B7-1 BINDING BY ANTI- PD-L1 NEUTRALIZING ANTIBODY
Serial dilutions of Anti- PD-L1 Neutralizing antibody (Catalog # EP134-C04) (1:1 serial dilution, from 10 μg/mL to 0.039μg/mL) was added into Biotinylated PD-L1: B7-1 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).