Revvity, 62H4TPAE, HTRF EPIgeneous Human and Mouse Total Histone H4 cellular Detection Kit, 500 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
Total Histone H4 assay principle
The Total Histone H4 assay measures Histone H4 levels in cells. Unlike Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis, or transfer. The assay uses 2 antibodies, one labeled with a donor fluorophore and the other with an acceptor. Both antibodies are highly specific for a distinct epitope on the protein.
In the presence of Histone H4, this enables an immune-complex formation involving both labeled antibodies, and which brings the donor fluorophore into close proximity to the acceptor, thereby generating a FRET signal. Its intensity is directly proportional to the concentration of total protein present in the sample.

Total 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 Total Histone H4 HTRF detection reagents. This protocol allows the cells' viability and confluence to be monitored.

Total Histone H4 one-plate assay protocol
Detection of Total 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-Total
Product Group-Kit
Sample Volume-10 µL
Shipping Conditions-Shipped in Dry Ice
Target-Histone H4
Target Class-Epigenetics
Target Species-Human Mouse
Technology-TR-FRET
Therapeutic Area-Immuno-oncology Oncology
Unit Size-500 assay points