The EpiQuik Global Mono-Methyl Histone H4-K20 Quantification Kit (Colorimetric) is a convenient package of tools that allows the experimenter to specifically measure global mono-methylation of histone H4-K20 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 with no need for radioactivity, electrophoresis, or chromatography.
- Specifically captures mono-methylated H4-K20 with the detection limit as low as 5 ng/well and detection range from 20 ng-2 µg/well of histone extracts.
- The control is conveniently included for the quantification of mono-methylated H4-K20.
- 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 transcriptional repression or activation. HMTs transfer 1-3 methyl groups from S-adenosyl-Lmethionine to the lysine and arginine residues of histone proteins. PR-SET7, SET9, SUV4.20h, and ASH1are histone methyltransferases that catalyze methylation of histone H4 at lysine 20 (H4-K20) in mammalian cells. H4-K20 mono-methylation is involved in the maintenance of the proper higher order structure of DNA and is consequently essential for chromosome condensation, it also functions in gene silencing. The global H4-K20 mono-methylation can also be changed by inhibition or activation of HMTs, making quantitative detection of global mono-methyl histone H4- K20 useful in developing a better understanding of epigenetic regulation of gene activation/repression and for developing HMT-targeted drugs. The EpiQuik™ Global Mono-Methyl Histone H4-K20 Quantification Kit (Colorimetric) provides a tool for measuring global mono-methylation of histone H4-K20.
Principle & Procedure
This kit is designed for measuring global histone H4-K20 mono-methylation. In an assay with this kit, the mono-methylated histone H4 at lysine 20 is captured to the strip wells coated with an anti-monomethyl H4-K20 antibody. The captured mono-methylated histone H4-K20 can then be detected with a detection antibody, followed by a color development reagent. The ratio of mono-methylated H4-K20 is proportional to the intensity of absorbance. The absolute amount of mono-methylated H4-K20 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.
C1 (10X Wash Buffer)
C2 (Antibody Buffer)
C3 (Detecting Antibody, 1 mg/ml)*
C4 (Color Developer)
C5 (Stop Solution)
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.