The EpiQuik™ Global Pan-Methyl Histone H3-K36 Quantification Kit (Colorimetric) is a convenient package of tools that allows the experimenter to specifically measure global mono-, di-, and tri-methylation of histone H3-K36 colorimetrically using a variety of mammalian cells (human, mouse, etc.) including fresh and frozen tissues, 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, and chromatography.
- Simultaneous quantification of mono-, di-, and tri-methylated H3-K36 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 mono-, di, and tri-methylated H3-K36.
- Strip microplate format makes the assay flexible: manual or high throughput.
- Simple, reliable, and consistent assay conditions.
Epigenetic activation or inactivation of genes plays a critical role in many important human diseases, especially in cancer. A major mechanism for epigenetic inactivation of the genes is methylation of CpG islands in genome DNA caused by DNA methyltransferases. Histone methyltransferases (HMTs) control or regulate DNA methylation through chromatin-dependent transcription repression or activation. HMTs transfer 1-3 methyl groups from S-adenosyl-Lmethionine to the lysine and arginine residues of histone proteins. SET2 is a histone methyltransferase that catalyzes methylation of histone H3 at lysine 36 (H3-K36) in mammalian cells. H3-K36 mono-methylation is commonly found together with di-methylation of H3-K27 and associated with transcriptionally silenced genes. H3-K36 di- and tri-methylation are associated with transcriptionally active genes. Increased global H3-K36 methylation is also found to be linked to the Sotos syndrome and leukemia-associated protein NSD1, and the Huntington disease protein HYPB. The patterns of global H3-K36 methylation can change by inhibition or activation of HMTs. Thus, quantitative detection of global mono, di-, and tri-methyl histone H3-K36 would provide useful information for better understanding epigenetic regulation of gene activation and for developing HMT-targeted drugs. The EpiQuik™ Global Pan-Methyl Histone H3-K36 Quantification Kit (Colorimetric) provides a tool for measuring global mono-, di-, and trimethylation of histone H3-K36.
Principle & Procedure
This kit is designed for measuring global histone H3-K36 mono-, di-, and tri-methylation. In an assay with this kit, the methylated histone H3 at lysine 36 is captured to the strip wells coated with antibodies specifically for mono-, di-, and tri-methyl H3-K36. The captured mono-, di-, and tri-methylated histone H3-K36 can then be detected with a labeled detection antibody, followed by a color development reagent. The ratio of mono-, di-, and tri-methylated H3-K36 is proportional to the intensity of absorbance. The absolute amount of the methylated H3-K36 can be quantitated 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.