ISK-012

PDCD1; PDCD1LG2

ID Components Size (96 tests)
ISK-012-1 High-bind Plate 1 plate
ISK-012-2 Human PD-L2 50 μg
ISK-012-3 Biotinylated Human PD-1 10 μg
ISK-012-4 Anti-PD-1 Neutralizing Antibody 20 μg
ISK-012-5 Streptavidin-HRP 10 μg
ISK-012-6 Coating Buffer 12 mL
ISK-012-7 10xWashing Buffer 50 mL
ISK-012-8 Blocking Buffer 50 mL
ISK-012-9 Substrate Solution 24 mL
ISK-012-10 Stop Solution 7 mL

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.

96 tests

5-0.01μg/mL

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.

This kit is developed for screening for inhibitors of human PD1 binding to human PD-L2

See Certificate of Analysis for details of reconstitution instruction and specific concentration.

The unopened kit is stable for 12 months from the date of manufacture if stored at 2°C to 8°C.
The opened kit should be stored per components table. The shelf life is 30 days from the date of opening.

Serial dilutions of Anti-PD-1 Neutralizing antibody (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.