膜杰作
Star Staining
| ID | Components | Size |
| EP120-C01 | High-bind Plate | 1 plate |
| EP120-C02 | Human PD-L2 | 50 μg |
| EP120-C03 | Biotinylated Human PD-1 | 10 μg |
| EP120-C04 | Anti-PD-1 Neutralizing Antibody | 20 μg |
| EP120-C05 | Streptavidin-HRP | 10 μg |
| EP120-C06 | Coating Buffer | 12 mL |
| EP120-C07 | 10xWashing Buffer | 50 mL |
| EP120-C08 | Blocking Buffer | 50 mL |
| EP120-C09 | Substrate Solution | 24 mL |
| EP120-C10 | Stop Solution | 7 mL |
背景（Background）
PD-L2, also known as CD273 or B7-DC, is a member of the B7 family protein with PD-L1, and it is the second important ligand that can bind pD-1 after PD-L1. The human PD-L2(B7-H2, CD273) gene is also located on chromosome 9p24, encoding type I transmembrane protein with 247 amino acids. The homology of PD-L1 and PD-L2 was 40%. The affinity between PD-L2 and PD-1 is 2-6 times that of PD-L1. Compared with PD-L1, the expression of PD-L2 is relatively limited, and pD-L2 is induced to express on the surface of dendritic cells, macrophages, bone marrow derived mast cells and other cells.
应用说明（Application）
This kit is developed for screening for inhibitors of human PD1 binding to human PD-L2.
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-1 pathway inhibitors. The assay takes advantage of our in house-developed binding of biotinylated human PD-1 to immobilized human PD-L2 in a functional ELISA assay, and employs a simple colorimetric ELISA platform. Briefly, we provide you with a human Biotinlated PD-1 protein, a human PD-L2 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-L2.
b) Add your molecule of interest to the tests.
c) Add human PD-1-Biotin to bind the coated human PD-L2.
d) Add Streptavidin-HRP followed by TMB or other colorimetric HRP substrate.
Finally, the half maximal inhibitory concentration (IC50) of your compound to PD-L2: PD-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-1 [BIOTINYLATED]: PD-L2 BINDING BY ANTI-PD-1 NEUTRALIZING ANTIBODY
Serial dilutions of Anti-PD-1 Neutralizing antibody (Catalog # EP120-C04) (1:1 serial dilution, from 5 μg/mL to 0.01μg/mL) was added into PD-L2: Biotinlated PD-1binding reactions. The assay was performed according to the protocol described above. Background was subtracted from data points prior to log transformation and curve fitting (QC tested).
