EpiQuik In Situ Histone H3 Phosphorylation (Ser10) Assay Kit

The EpiQuik™ In Situ Histone H3 Phosphorylation (Ser10) Assay Kit is a convenient set of tools that allows the experimenter to measure in situ phospho histone H3 (ser10). The kit is ready-to-use and provides all the essential components needed for specifically measuring histone H3 phosphorylation (ser10) in situ using cultured adherent cells. The kit has the following advantages:

• Quick and efficient procedure, which can be finished within 3 hours.
• Innovative colorimetric assay without the need for radioactivity, electrophoresis, or chromatography.
• Measurement of in situ histone H3 phosphorylation at ser10 and without the need to prepare cell lysates.
• Microplate format makes the assay suitable for high throughput analysis of reagents that increases or inhibits histone H3 phosphorylation at ser10.
• Simple, reliable, and consistent assay conditions.

Principle & Procedure
EpiQuik™ In Situ Histone H3 Phosphorylation (Ser10) Assay Kit is a whole cell-based detection kit of histone H3 phosphorylation (ser10). In this assay, adherent cells are cultured in conventional 96-well microplates. After your experimental treatment, cells are fixed and permeabilized. The histone H3 phosphorylation (ser10) is then detected by an anti-phospho histone H3 (ser10) antibody. The ratio or amount of phospho histone H3 (ser10) can be quantified through HRP conjugated secondary antibody-color development system and is proportional to the intensity of color development.

 
Fig. 1. Schematic procedure for using the EpiQuik In Situ Histone H3 Phosphorylation (Ser10) Assay Kit.

Fig. 2. Log-phase MCF-7 cells were serum-starved overnight, and serum was added back for 30 minutes followed by treatment with Calyculin A (CL-A, 0.2 μM for 30 minutes). The level of H3 phosphorylation at ser10 was measured with this kit.