EpiQuik Circulating Total Histone H3 Quantification Kit (Colorimetric)

The EpiQuik Circulating Total Histone H3 Quantification Kit (Colorimetric) is a complete set of optimized buffers and reagents for specifically designed for quantifying levels of histone H3 proteins independent of its modified state and can also be used for normalizing the modified histone H3 content of samples when run in parallel with Epigentek circulating modified histone H3 ELISA kit series. The kit has the following features:

• Quick and efficient procedure, which can be finished within 2.5 hours.
• High sensitivity and specificity. The detection limit is as low as 2 ng/well with dynamic range of 5-200 ng/well within the indicated amount range of the plasma/serum.
• The control is conveniently included for the quantification of H3.
• Strip microplate format makes the assay flexible: manual or high throughput.
• Simple, reliable, and consistent assay conditions.

Background Information
Histone H3, along with H2A, H2B, and H4, is involved in the structure of chromatin in eukaryotic cells. Histone H3 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, phosphorylation, methylation, ubiquitination, and ADP-ribosylation, occur on the N-terminal tail domains of histone H3 through catalyzing of histone modifying enzymes, which result in remodeling of the nucleosome structure into an open conformation more accessible to transcription complexes. In most species, histone H3 is primarily acetylated at lysine 9, 14, 18, 23, and 56; methylated at lysine 4, 9, 27, 36, and 79; and phosphorylated at ser10, ser28, Thr3, and Thr11, respectively.  Recently, H3 citrullination was also observed and related to several disease or pathological status. Thus, quantitative detection of various histone modifications, especially from plasma or serum samples  would provide useful information for better understanding epigenetic regulation of cellular processes and for discovering new circulating biomarkers. 

Principle & Procedure
This kit is designed for measuring total histone H3 in plasma or serum. In an assay with this kit, the histone H3 proteins in the plasma/serum sample are captured on the strip wells coated with anti-histone H3 antibody. The captured histoen H3 can then be recognized with detection antibody followed by a color development reagent. The ratio of histone H3 is proportional to the intensity of absorbance. The absolute amount of H3 can be quantitated by comparing to the standard control.

Starting Materials
Input materials should be plasma or serum. The amount of plasma or serum for each assay can be 10 to 40 µl with an optimal amount of 30 µl.