The EpiQuik™ Global Acetyl Histone H3-K9 Quantification Kit (Fluorometric) is a convenient package of tools that allows the experimenter to specifically measure global acetylation of histone H3-K9 fluorometrically, 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 fluorometric assay without the need for radioactivity, electrophoresis, or chromatography.
- Specifically captures acetyl H3-K9 with the detection limit as low as 0.4 ng/well and detection range from 5 ng-2 µg/well of histone extracts.
- The control is conveniently included for the quantification of the amount of acetyl H3-K9.
- Strip microplate format makes the assay flexible: manual or high throughput.
- Simple, reliable, and consistent assay conditions.
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 (HAT) and histone deacetylases (HDACs) play a critical role in control of histone H3 acetylation at multiple sites. Histone H3 at lysine 9 (H3-K9) is a primary acetylated site of histone H3. Acetylation of histone H3-K9 appears to have a dominant role in histone deposition and chromatin assembly in some organisms. Acetylation of histone H3-K9 is tightly involved in the cell cycle regulation, cell proliferation, and apoptosis. Acetylation of histone H3-K9 is also an active marker, and the balance between histone H3-K9 acetylation and methylation is important for the establishment of specific chromatin structures. An imbalance in the equilibrium of histone H3 acetylation, including K9 acetylation, has been associated with tumorigenesis and cancer progression. Histone H3-K9 acetylation may be increased by inhibition of HDACs and decreased by HAT inhibition. Thus, quantitative detection of global acetyl histone H3-K9 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 H3-K9 Quantification Kit (Fluorometric) provides a tool for measuring global acetylation of histone H3-K9.
Principle & Procedure
This kit is designed for measuring global histone H3-K9 acetylation. In an assay with this kit, the acetyl histone H3 at lysine 9 is captured to the strip wells coated with an anti-acetyl H3-K9 antibody. The captured acetyl histone H3-K9 can then be detected with a labeled detection antibody followed by a fluorescent development reagent. The ratio of acetyl H3-K9 is proportional to the intensity of fluorescence. The absolute amount of acetyl H3-K9 can be quantified by comparing to the standard control.
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.
Fig. 1. Schematic procedure for using the EpiQuik Global Acetyl Histone H3K9 Quantification Kit (Fluorometric).