Revvity, 62NH3PAD, HTRF Total Histone H3 Detection Kit, 10,000 Assay Points

Overview
This cellular assay is for the direct detection of endogenous levels of Total H3 (histone 3) in cells. The assay can be used for adherent cells or suspension cells, primary or secondary screening, inhibitor studies and normalization studies of both methyltransferases and demethylases. The total H3 kit has been validated with Revvity's EPIgeneous H3K27ME3 and H3K36ME2 kits for normalization studies and be applied to the same assay lysates.
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
This assay is designed for the simple, rapid and direct detection of endogenous levels of H3 Total in cells. Histone H3 is detected in a sandwich assay format using 2 different specific antibodies, one labelled with Eu-cryptate (donor) and the second with d2 (acceptor). The HTRF signal is proportional to the concentration of histone H3.

 

Two-plate assay protocol
Cells are plated, (stimulated) and lysed in the same culture plate and then transferred to the assay plate for the detection of Total H3 by HTRF reagents. This protocol allows the cells' viability and confluence to be monitored. Moreover, the Total H3 kit has been designed to be run in parallel with our EPIgeneous specific methyl mark cellular assay. With this, a high lysis volume makes it possible to assess both methylation marks and total H3 from the same 96w lysate.

 

One-plate assay protocol
Detection of Total H3 with HTRF reagents is performed in a single plate used for plating, stimulation and detection. No washing steps are required. This protocol, HTS designed, allows miniaturization while maintaining HTRF quality. Application-Biochemical Enzymatic Assay
Brand-HTRF
Detection Modality-HTRF
Molecular Modification-Total
Product Group-Kit
Sample Volume-16 µL
Shipping Conditions-Shipped in Dry Ice
Target Class-Epigenetics
Technology-TR-FRET
Therapeutic Area-Metabolism/Diabetes Neuroscience Oncology & Inflammation
Unit Size-10,000 assay points