Revvity, 62H4PANACPAD, HTRF EPIgeneous Human and Mouse Pan-Acetyl Histone H4 cellular Detection Kit, 10,000 assay points

Overview
Histone 4 is one of the core histone proteins involved - along with other histones - in the packaging of DNA into a condensed chromatin state. Histone H4 function is dependent on its acetylation state (1 acetyl group per lysine) at several sites (Lys5, Lys8, Lys12, and Lys16) on the N-terminal tail. The mis-regulation of histone acetylation is closely associated with cancer initiation and progression, making histone acetylase and deacetylase enzymes promising drug targets.
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 readout time or size of assays
How it works
Pan-Acetyl Histone H4 assay principle
The Pan-Acetyl Histone H4 assay measures Histone H4 when acetylated at Lys5, Lys8, Lys12, and Lys16. Unlike Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis, or transfer. The acetylation on Lysines 5, 8, 12, and 16 of histone H4 is detected in a sandwich assay format using 2 different specific antibodies, one labelled 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 (laser or flash lamp) triggers a Fluorescence Resonance Energy Transfer (FRET) towards the acceptor, which in turn fluoresces at a specific wavelength (665 nm). One reagent binds to Histone H4 and the other binds to acetyl K5-K8-K12-K16 Histone H4, thereby generating a FRET signal. Its intensity is directly proportional to the concentration of acetylated protein present in the sample and provides a means of assessing the protein’s acetylation state under a no-wash assay format.

Pan-Acetyl Histone H4 two-plate assay protocol
The two-plate protocol involves culturing cells in a 96-well plate before lysis, then transferring lysates into a 384-well low volume detection plate before the addition of Pan-Acetyl Histone H4 HTRF detection reagents. This protocol allows the cells' viability and confluence to be monitored.

Pan-Acetyl Histone H4 one-plate assay protocol
Detection of Phosphorylated Pan-Acetyl Histone H4 with HTRF reagents can be performed in a single plate used for culturing, stimulation, and lysis. No washing steps are required. This HTS designed protocol allows miniaturization while maintaining robust HTRF quality. Application-Cell Signaling
Brand-HTRF
Detection Modality-HTRF
Lysis Buffer Compatibility-EPIgeneous Lysis Buffer A
Molecular Modification-Acetylation
Product Group-Kit
Sample Volume-10 µL
Shipping Conditions-Shipped in Dry Ice
Target-Pan-Acetyl Histone H4
Target Class-Epigenetics
Target Species-Human Mouse
Technology-TR-FRET
Therapeutic Area-Immuno-oncology Oncology
Unit Size-10,000 assay points