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Your Position: Home > Non SARS-Cov-2 Kits > PD-1 > EP-120

PD-1 [Biotinylated] : PD-L2 Inhibitor Screening ELISA Kit

For research use only.
IDComponentsSize
EP120-C01High-bind Plate1 plate
EP120-C02Human PD-L250 μg
EP120-C03Biotinylated Human PD-110 μg
EP120-C04Anti-PD-1 Neutralizing Antibody20 μg
EP120-C05Streptavidin-HRP10 μg
EP120-C06Coating Buffer12 mL
EP120-C0710xWashing Buffer 50 mL
EP120-C08Blocking Buffer50 mL
EP120-C09Substrate Solution24 mL
EP120-C10Stop Solution7 mL
  • 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.
  • Application
    This kit is developed for screening for inhibitors of human PD1 binding to human PD-L2

    It is for research use only.

  • Reconstitution
    Please see Certificate of Analysis for details of reconstitution instruction and specific concentration.
  • Storage
    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.

  • 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.

Typical Data
 PD-1 ELISA

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).

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