EpiQuik Global Acetyl Histone H3K27 Quantification Kit (Colorimetric)

The EpiQuik™ Global Acetyl Histone H3K27 Quantification Kit (Colorimetric) is a convenient package of tools that allows the experimenter to specifically measure global acetylation of histone H3-K27 colorimetrically, using a variety of mammalian cells (human, mouse, etc.) including fresh and frozen tissues, and cultured adherent and suspension cells. The kit has the following advantages:

• Quick and efficient procedure, which can be finished within 2.5 hours.
• Innovative colorimetric assay without the need for radioactivity, electrophoresis, or chromatography.
• Specifically captures acetyl H3-K27 with the detection limit as low as 2 ng/well and detection range from 20 ng-1 µg/well of histone extracts.
• The control is conveniently included for the quantification of the amount of acetyl H3-K27.
• Strip microplate format makes the assay flexible: manual or high throughput.
• Simple, reliable, and consistent assay conditions.

Background Information
Acetylation of histones, including histone H3, has been involved in the regulation of chromatin structure and recruitment of transcription factors to the gene promoters. Histone acetyltransferases (HATs) and histone deacetylases (HDACs) play a critical role in the control of histone H3 acetylation at multiple sites. Histone H3 at lysine 27 (H3-K27) is one of the primary acetylated sites of histone H3. Acetylation of histone H3-K27 is tightly involved in cell cycle regulation, cell proliferation, and apoptosis; acetylation of histone H3-K27 is also an active marker. For example, loss of Polycomb Repressive Complex 2 (PRC2) activity results in a global increase in H3K27 acetylation suggesting that PRC2 represses transcription by preventing the binding of acetyltrasferases to K27 position. H3-K27 acetylation shows a robust peak at the transcription start site (TSS) of active and poised genes. The balance between histone H3-K27 acetylation and methylation is important for the establishment of specific chromatin structures. An imbalance in the equilibrium of histone H3 acetylation, including K27 acetylation, has been associated with tumorigenesis and cancer progression. Histone H3-K27 acetylation may be increased by inhibition of HDACs and decreased by HAT inhibition. Thus, quantitative detection of global acetyl histone H3-K27 would provide useful information for better understanding epigenetic regulation of gene activation and for developing HAT or HDAC-targeted drugs. The EpiQuik™ Global Acetyl Histone H3K27 Quantification Kit (Colorimetric) provides a tool for measuring global acetylation of histone H3-K27.

Principle & Procedure
This kit is designed for measuring global histone H3-K27 acetylation. In an assay with this kit, the acetyl histone H3 at lysine 27 is captured to the strip wells coated with an anti-acetyl H3-K27 antibody. The captured acetyl histone H3-K27 can then be detected with a labeled detection antibody, followed by a color development reagent. The ratio of acetyl H3-K27 is proportional to the intensity of absorbance. The absolute amount of acetyl H3-K27 can be quantified by comparing to the standard control.

Starting Materials
Input material should be purified histone extracts. In general, the input amount should be from 50 ng to 200 ng per well of histone extracts.