Revvity, 64SIRPPEG, HTRF Human CD47 / SIRP-α Binding Kit, 500 Assay Points

Overview
Signal-regulatory protein alpha (SIRP-alpha), also known as CD172a, is a cell surface protein expressed mainly by myeloid cells, such as macrophages. Its receptor, CD47, is ubiquitously expressed on the surface of all cells and has been found to be overexpressed in some cancers. CD47 binding to SIRP-alpha delivers a "Do Not Eat Me" signal to macrophages. In the case of a tumor, this signaling aids in tumor evasion of the immune system. Inhibitors of this interaction hold great promise for cancer immunotherapy.
HTRF assays offer many advantages over other technologies:

Homogeneous add-and-read format
No wash steps
Low background
Straightforward miniaturization from 96- or 384-well microplates to high density assay formats such as 384-well low volume and 1536-well plates
Stable signal, providing flexibility in time of readout or size of assays
How it works
Assay principle
The CD47/SIRP binding assay includes human recombinant immune checkpoint partners tagged for detection (CD47 and SIRP alpha) and labelled anti-tag reagents for HTRF detection. Without an inhibitor, CD47 binds to SIRP alpha, and the binding of each detection reagent to its tagged target generates an HTRF signal. In the presence of compound, standard or an antibody blocking the CD47/SIRP alpha interaction, the HTRF signal decreases.

 

Assay protocol
This CD47/SIRP alpha binding assay can be run in a 96- or 384-well low volume white plate (20 µL final). As described here, samples or standards are dispensed directly into the assay plate, and the tagged CD47 & SIRP alpha protein are then added, followed by the dispensing of the HTRF reagents. The reagents labelled with HTRF fluorophores may be pre-mixed and added in a single dispensing step. No washing steps are needed. The protocol can be further miniaturized or upscaled by simply resizing each addition volume proportionally. Application-Protein-Protein Interaction
Brand-HTRF
Detection Modality-HTRF
Product Group-Kit
Sample Volume-2 µL
Shipping Conditions-Shipped in Dry Ice
Target Class-Binding Assay
Target Species-Human
Technology-TR-FRET
Therapeutic Area-Oncology & Inflammation
Unit Size-500 assay points