ID | Components | Size |
EP152-C01 | High-bind Plate | 1 plate |
EP152-C02 | Human SIRP alphaV2 | 20 μg |
EP152-C03 | Anti-CD47 Neutralizing Antibody | 20 μg |
EP152-C04 | Human CD47-Biotin | 10 μg |
EP152-C05 | Streptavidin-HRP | 10 μg |
EP152-C06 | Coating Buffer | 12 mL |
EP152-C07 | 10xWashing Buffer | 50 mL |
EP152-C08 | Blocking Buffer | 50 mL |
EP152-C09 | Substrate Solution | 12 mL |
EP152-C10 | Stop Solution | 7 mL |
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-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.
应用说明(Application)
This kit is developed for screening for inhibitors of human CD47 binding to human SIRP alphaV2.
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)
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.
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 alphaV2 in a functional ELISA assay, and employs a simple colorimetric sandwich ELISA platform.
Your experiment will include 4 simple steps:
a) Coat the plate with human SIRP alphaV2.
b) Add your molecule of interest to the tests.
c) Add human CD47-Biotin to bind the coated human SIRP alphaV2.
d) Add Streptavidin-HRP followed by TMB or other colorimetric HRP substrate.
Finally, the ability of your compound to inhibit CD47 [Biotinylated] : SIRP alphaV2 Rbinding 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 ALPHAV2 BINDING BY ANTI-CD47 NEUTRALIZING ANTIBODY
Serial dilutions of Anti-CD47 Neutralizing Antibody (Catalog # EP152-C03) (1:1 serial dilution, from 10 μg/mL to 0.078 μg/mL (68.493-0.535 nM)) was added into SIRP alphaV2: CD47-Biotin binding reactions. The assay was performed according to the above-described protocol. Background was subtracted from data points prior to log transformation and curve fitting (QC tested).