EpiQuik Global Acetyl Histone H4K5 Quantification Kit (Colorimetric)

The EpiQuik™ Global Acetyl Histone H4-K5 Quantification Kit (Colorimetric) is a convenient package of tools that allows the experimenter to specifically measure global acetylation of histone H4-K5 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 H4-K5 with the detection limit as low as 2 ng/well and detection range from 20 ng-5 µg/well of histone extracts.
• The control is conveniently included for the quantification of the amount of acetyl H4-K5.
• 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 and H4, have 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 H4 acetylation at multiple sites. Acetylation of histone H4 at lysine 5 (H4-K5) reflects the hyperacetylated state in histone H4 and is strongly correlated with active states of genes. H4-K5 acetylation is considered to be a marker of dexamethasone transactivation and the change of H4- K5 acetylation is observed in cancer and inflammatory diseases. Histone H4-K5 acetylation may be increased by inhibition of HDACs and decreased by HAT inhibition; thus, quantitative detection of global acetyl histone H4-K5 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 H4-K5 Quantification Kit (Colorimetric) provides a tool for measuring global acetylation of histone H4-K5.

Principle & Procedure
This kit is designed for measuring global histone H4-K5 acetylation. In an assay with this kit, the acetyl histone H4 at lysine 5 is captured to the strip wells coated with an anti-acetyl H4-K8 antibody. The captured acetyl histone H4-K5 can then be detected with a labeled detection antibody followed by a color development reagent. The ratio of acetyl H4-K5 is proportional to the intensity of absorbance. The absolute amount of acetyl H4-K5 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.