Revvity, 62KA2PAD, HTRF EPIgeneous 2-methyl K4 Histone H3 Detection Kit, 10,000 Assay Points

Overview
This cellular assay has been developed with optimized reagents and protocols for the direct detection of endogenous levels of H3K4Me2.
This HTRF® format uses a very specific antibody pair combined with a highly efficient nucleus extraction which enables the assay to be used with a variety of cell backgrounds, and for the study of important methyltransferases and demethylases targets, such as JARID1 and MLL1.
The H3K27Me3 assay can be used for adherent or suspension cells, primary or secondary screening, and inhibitor studies.
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 dimethylation of Lysine 4 on histone H3 is detected in a sandwich assay format using 2 different specific antibodies, one labeled with Eu-Cryptate (donor) and the second with d2 (acceptor). When the dyes are in close proximity, the excitation of the donor with a light source triggers a Fluorescence Resonance Energy Transfer (FRET), which in turn fluoresces at a specific wavelength (665 nm). One conjugate binds to Histone H3 and the other binds to the K4Me2 mark. The specific signal modulates positively in proportion to trimethylation on Lysine 4.

 

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 H3K4Me2 by HTRF reagents. This protocol enables the cells viability and confluence to be monitored.

 

One-plate assay protocol
Detection of H3K4Me2 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-EPIgeneous
Detection Modality-HTRF
Molecular Modification-Methylation
Product Group-Kit
Sample Volume-120 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