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Your Position: Home > Kits > CD47 > EP-160

CD47 : SIRP gamma [Biotinylated] Inhibitor Screening ELISA Kit

For research use only.

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EP160-C01High-bind Plate1 plate
EP160-C02Human CD4735 μg
EP160-C03Biotinylated Human SIRP gamma15 μg
EP160-C04Anti-CD47 Neutralizing Antibody20 μg
EP160-C05Streptavidin-HRP5 μg
EP160-C06Coating Buffer12 mL
EP160-C0720xWashing Buffer 50 mL
EP160-C08Blocking Buffer50 mL
EP160-C09Substrate Solution12 mL
EP160-C10Stop Solution7 mL
  • Background
    CD47 is a transmembrane protein, a cell surface glycoprotein molecule belonging to the immunoglobulin superfamily that binds to a variety of proteins, including integrins, thrombospondin 1, and signal regulatory protein-α (SIRPα). In recent years, accumulating data have demonstrated that the CD47-SIRPαV2 axis is a key immune checkpoint in different cancers, including hematological malignancies, similar to PD-1/PD-L1 in solid tumors. CD47-SIRPαV2 blockade has become a next-generation immune checkpoint blocking strategy for various malignancies after PD-1/PD-L1.
  • Application

    This kit is developed for screening for inhibitors of human CD47 binding to human SIRP alphaV2.

    It is for research use only.

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

  • Assay Principles
    This inhibitor screening ELISA kit is designed to facilitate the identification and characterization of new CD47 pathway inhibitors. The assay takes advantage of our in house-developed binding of biotinylated human SIRP gamma to immobilized human CD47 in a functional ELISA assay, and employs a simple colorimetric ELISA platform. Briefly, we provide you with a human Biotinylated SIRP gamma protein, a human CD47 protein, an anti-CD47 neutralizing antibody (as method verified Std.), and Streptavidin-HRP reagent.

    Your experiment will include 4 simple steps:

    a) Coat the plate with human CD47.

    b) Add your molecule of interest to the tests.

    c) Add human SIRP gamma-Biotin to bind the coated CD47.

    d) Add Streptavidin-HRP followed by TMB or other colorimetric HRP substrate.

    Finally, the half maximal inhibitory concentration (IC50) of your compound to CD47:SIRP gamma binding will be determined by comparing OD readings among different experimental groups.

Typical Data Please refer to Ds document for the assay protocol.

CD47: SIRP gamma [Biotinylated] Inhibitor Screening ELISA Kit
Serial dilutions of Anti-CD47 Neutralizing antibody (Catalog # EP160-C04) (1:1 serial dilution, from 5μg/mL to 0.039μg/mL) was added into CD47: SIRP gamma [Biotinylated] 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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