The EpiQuik™ Global Acetyl Histone H4-K12 Quantification Kit (Fluorometric) is a convenient package of tools that allows the experimenter to fluorometrically measure global acetylation of histone H4-K12. The kit is ready-to-use and provides all the essential components needed to carry out a successful assay experiment and is suitable for specifically measuring global histone H4-K12 acetylation 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 H4-K12 with the detection limit as low as 0.4 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-K12.
- Strip microplate format makes the assay flexible: manual or high throughput.
- Simple, reliable, and consistent assay conditions.
Acetylation of histones, including histone H3 and H4, have been involved in the regulation of chromatin structure and the 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 12 (H4-K12) reflects the hyperacetylated state in histone H4 and is strongly correlated to active states of genes. H4-K12 acetylation is observed in cancer and inflammatory diseases. Histone H4-K12 acetylation may be increased by the inhibition of HDACs and decreased by HAT inhibition. Thus, quantitative detection of global acetyl histone H4-K12 would provide useful information for better understanding epigenetic regulation of gene activation, and for developing HAT or HDACtargeted drugs. The EpiQuik™ Global Acetyl Histone H4-K12 Quantification Kit (Fluorometric) provides a tool for measuring global acetylation of histone H4-K12.
Principle & Procedure
This kit is designed for measuring global histone H4-K12 acetylation. In an assay with this kit, the acetyl histone H4 at lysine 12 is captured to strip wells coated with an acetyl H4-K12 antibody. The captured acetyl histone H4-K12 can then be detected with a labeled detection antibody followed by a fluorescent development reagent. The ratio of acetyl H4-K12 is proportional to the intensity of fluorescence. The absolute amount of acetyl H4-K12 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.
F1 (10X Wash Buffer)
F2 (Antibody Buffer)
F3 (Detection Antibody, 1 mg/ml)*
F4 (Fluoro Developer)*
F5 (Fluoro Enhancer)*
F6 (Fluoro Dilution)
Standard Control (100 µg/ml)*
Signal Report Solution*
8-Well Sample Strips (with Frame)
8-Well Standard Control Strips
* For maximum recovery of the products, centrifuge the original vial prior to opening the cap.