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Your Position: Home > Kits > PD-1 > EP-154

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

For research use only.
IDComponentsSize
EP154-C01High-bind Plate1 plate
EP154-C02Human PD-115 μg
EP154-C03Biotinylated Human PD-L220 μg
EP154-C04Anti-PD1 Neutralizing Antibody20 μg
EP154-C05Streptavidin-HRP5 μg
EP154-C06Coating Buffer12 mL
EP154-C0710xWashing Buffer 50 mL
EP154-C08Blocking Buffer50 mL
EP154-C09Substrate Solution12 mL
EP154-C10Stop Solution7 mL
  • 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.
  • Application

    This kit is developed for screening for inhibitors of human PD-1 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-L2 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-L2 protein, a human PD-1 protein, an anti-PD1 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-L2-Biotin to bind the coated 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-L2 binding will be determined by comparing OD readings among different experimental groups.

Typical Data Please refer to DS document for The assay protocol.
PD-1 TYPICAL DATA

INHIBITION OF PD-1: PD-L2 [Biotinylated] BINDING BY ANTI-PD-1 NEUTRALIZING ANTIBODY
Serial dilutions of Anti-PD-1 Neutralizing antibody (Catalog # EP154-C04) (1:1 serial dilution, from 5 μg/mL to 0.0098μg/mL) was added into PD-1: Biotinylated PD-L2 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).

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