Revvity, 64HYGSTPEG, HTRF Hybridoma (GST-antigen) Binding Kit, 500 Assay Points

Overview
The hybridoma technology is still the most common technique used to generate antibodies for therapeutic applications. The generation and screening of hybridomas from an immunized animal involve a time-consuming process and sample only a fraction of the antibodies generated during the adaptive immune response. Therefore, screening hybridoma supernatants for their ability to efficiciently bind the target of interest has become crucial.
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 mouse hybridoma supernatant binding to a GST-tagged antigen is measured using a sandwich immunoassay, involving an anti-GST antibody labeled with an HTRF donor and an anti-mouse antibody labeled with an acceptor dye. The binding of murine antibodies produced by the hybridoma clone to their GST-tagged target generates an HTRF Signal.

 

Assay Protocol
The HTRF hybridoma binding kit can be run in a 96- or 384-well low volume white plate (20 µL final). As described here, samples are dispensed directly into the assay plate. The GST antigen is then added, followed by the dispensing of the HTRF reagents: the Anti GST antibody labeled with europium cryptate and the anti mouse antibody labeled with d2. 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-5 µL
Shipping Conditions-Shipped in Dry Ice
Target Class-Binding Assay
Technology-TR-FRET
Therapeutic Area-Inflammation Neuroscience Oncology & Inflammation
Unit Size-500 assay points