ID | Components | Size |
EP138-C01 | High-bind Plate | 1plate |
EP138-C02 | Human SIRP alpha | 15μg |
EP138-C03 | Biotinylated Human CD47 | 10μg |
EP138-C04 | Anti-CD47 Neutralizing Antibody | 20μg |
EP138-C05 | Streptavidin-HRP | 10μg |
EP138-C06 | Coating Buffer | 12mL |
EP138-C07 | 10xWashing Buffer | 50mL |
EP138-C08 | Blocking Buffer | 50mL |
EP138-C09 | Substrate Solution | 12mL |
EP138-C10 | Stop Solution | 12mL |
背景(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-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.
应用说明(Application)
This kit is developed for screening for inhibitors of human CD47 binding to human SIRP alpha.
It is for research use only.
重构方法(Reconstitution)
Please see Certificate of Analysis for details of reconstitution instruction and specific concentration.
存储(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.
原理(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 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.
典型数据-Typical Data Please refer to Ds document for the assay protocol.
INHIBITION OF CD47[Biotinylated]: SIRP alpha BINDING BY ANTI-CD47 NEUTRALIZING ANTIBODY
Serial dilutions of Anti-CD47 Neutralizing antibody (Catalog # EP138-C04) (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 (QC tested).