ISK-007

CD47; SIRPA

ID Components Size (96 tests)
ISK-007-1 High-bind Plate 1plate
ISK-007-2 Human SIRP alpha 15μg
ISK-007-3 Biotinylated Human CD47 10μg
ISK-007-4 Anti-CD47 Neutralizing Antibody 20μg
ISK-007-5 Streptavidin-HRP 10μg
ISK-007-6 Coating Buffer 12mL
ISK-007-7 10xWashing Buffer 50mL
ISK-007-8 Blocking Buffer 50mL
ISK-007-9 Substrate Solution 12mL
ISK-007-10 Stop Solution 12mL

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-SIRP-α axis is a key immune checkpoint in different cancers, including hematological malignancies, similar to PD-1/PD-L1 in solid tumors. Cd47-sirpα blockade has become a next-generation immune checkpoint blocking strategy for various malignancies after PD-1/PD-L1.

96 tests

10-0.01μg/mL

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 CD47 to immobilized human SIRP alpha in a functional ELISA assay, and employs a simple colorimetric ELISA platform. Briefly, we provide you with a human Biotinylated CD47 protein, a human SIRP alpha 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 SIRP alpha.
b) Add your molecule of interest to the tests.
c) Add human CD47-Biotin to bind the coated SIRP alpha.
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 alpha binding will be determined by comparing OD readings among different experimental groups.

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

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-CD47 Neutralizing antibody (1:1 serial dilution, from 10 μg/mL to 0.01μg/mL) was added into Biotinylated CD47: SIRP alpha 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.