EpiQuik Total Histone H4 Quantification Kit (Fluorometric)

The EpiQuik™ Total Histone H4 Quantification Kit (Fluorometric) is a complete set of essential components which enables an experimenter to fluorometrically quantify levels of histone H4 proteins independent of their modified states. It can also be used for normalizing the modified histone H4 content of samples when run in parallel with Epigentek's histone modification quantification kit series. The EpiQuik™ Total Histone H4 Quantification Kit (Fluorometric) is suitable for specifically measuring total histone H4 using human, mouse, and rat samples, including fresh and frozen tissues and cultured adherent and suspension cells.

• Quick and efficient procedure, which can be finished within 3 hours.
• Innovative fluorometric assay without the need for radioactivity, electrophoresis, or chromatography.
• Specifically captures histone H4 with the detection limit as low as 20 ng/well and detection range from 50 ng to 500 ng/well of histone extracts.
• The unmodified histone H4 control is conveniently included for the quantification of the amount of total histone H4.
• Strip microplate format makes the assay flexible: manual or high throughput.
• Simple, reliable, and consistent assay conditions.

Background Information
Histone H4, along with H2A, H2B, and H3 is involved in the structure of chromatin in eukaryotic cells. Histone H4 can undergo several different types of epigenetic modifications that influence cellular processes such as transcription activation/inactivation, chromosome packaging, and DNA damage/repair. These modifications, including acetylation and methylation, occur on the N-terminal tail domains of histone H4 through catalyzation of histone modifying enzymes. This results in the remodeling of the nucleosome structure into an open conformation which is more accessible to transcription complexes. Thus, quantitative detection of various histone modifications would provide useful information for better understanding epigenetic regulation of cellular processes and for developing HMT-targeted drugs.

Principle & Procedure
In an assay with this kit, the histone H4 protein is captured to strip wells coated with an anti-histone H4 antibody. The captured histone H4 can then be detected with a detection antibody, followed by a fluorescence development reagent. The ratio of histone H4 is proportional to the intensity of fluorescence by reading the relative fluorescence units (RFU) in a fluorescence microplate reader at 530ex/590em. The absolute amount of histone H4 can be quantified by comparing to the standard control.

Starting Materials
Input materials can be histone extracts or nuclear extracts. The amount of histone extracts for each assay can be 50 ng to 500 ng with an optimal range of 0.1 to 0.2 µg.