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 & ProcedureEpiQuik™ 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.