Revvity, 64HH3PEH, HTRF Human and Mouse Phospho-Histone H3 (Ser10) Detection Kit, 10,000 Assay Points

Overview
The Phospho-Histone H3 (Ser10) assay enables the detection of Ser10 phosphorylation in the tails of Histone H3, a key regulator of gene expression and mitosis. Phosphorylation at Ser10 is tightly correlated with chromosome condensation during both mitosis and meiosis.
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
Phospho-Histone H3 (Ser10) assay principle
The phospho-Histone H3 (Ser10) assay measures Histone H3 when phosphorylated at Ser10. Contrary to Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis or transfer. The phospho-Histone H3 (Ser10) assay uses 2 labeled antibodies: one with a donor fluorophore, the other one with an acceptor. The first antibody is selected for its specific binding to the phosphorylated motif on the protein, the second for its ability to recognize the protein independent of its phosphorylation state. Protein phosphorylation 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 phosphorylated protein present in the sample, and provides a means of assessing the proteins phosphorylation state under a no-wash assay format.

 

Phospho-Histone H3 (Ser10) 2-plate assay protocol
The 2 plate protocol involves culturing cells in a 96-well plate before lysis then transferring lysates to a 384-well low volume detection plate before adding Phospho-Histone H3 (Ser10) HTRF detection reagents. This protocol enables the cells' viability and confluence to be monitored.

 

Phospho-Histone H3 (Ser10) 1-plate assay protocol
Detection of Phosphorylated Histone H3 (Ser10) 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 enables miniaturization while maintaining robust HTRF quality. Application-Cell Signaling
Brand-HTRF
Detection Modality-HTRF
Molecular Modification-Phosphorylation
Product Group-Kit
Sample Volume-16 µL
Shipping Conditions-Shipped in Dry Ice
Target Class-Phosphoproteins
Target Species-Human Mouse
Technology-TR-FRET
Therapeutic Area-Neuroscience
Unit Size-10,000 assay points