|Product Overview:||Pan-Methyl Histone H3-K79 Quantification Kit (Fluorometric) is use for measuring mono, di-and tri-methylation of histone H3-K79.|
|Description:||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-L-methionine to the lysine and arginine residues of histone proteins. Dot1 is a histone methyltransferase that catalyzes methylation of histone H3 at lysine 79 (H3-K79) in mammalian cells. Intergenic H3-K79 mono-methylation is a mark of enhancer regions and may provide unique binding surfaces along the chromatin fiber to stabilize interactions between enhancers and their target promoters. H3-K79 diand tri-methylation is also a widespread histone modification and is associated with transcriptionally active genes. Increased H3-K79 methylation is found to be involved in some pathological processes such as leukemogenesis in human. The patterns of H3-K79 methylation can also be changed by inhibition or activation of HMTs. Thus quantitative detection of mono-, di- and tri-methyl histone H3-K79 would provide useful information for better understanding epigenetic regulation of gene activation and for developing HMT targeted drugs. The Pan-Methyl Histone H3-K79 Quantification Kit (Colorimetric) provides a tool for measuring mono, di- and tri-methylation of histone H3-K79.|
|Applications:||For simultaneously measuring histone H3-K79 mono-, di-, and tri-methylation using a variety of mammalian cells (human, mouse, etc.) including fresh and frozen tissues, cultured adherent and suspension cells.|
|Usage:||For research use only (RUO)|
|Storage:||Upon receipt, store F3, F4 and Standard control at –20°C. Store all other components at 4°C away from light.The components of the kit should be stable for 6 months when stored properly.Note: Check if buffers F1 and F2 contain salt precipitates before using. If so, warm (at room temperature or 37°C) and shake the buffers until the salts are redissolved.|
|Kit Components:||F1 (10X wash buffer) 20 mlF2 (antibody buffer) 12 mlF3 (detection antibody, 1 mg/ml)* 10 µlF4 (fluoro-developer)* 24 µlF5 (fluoro enhancer)* 24 µlF6 (fluoro-dilution) 8 mlStandard control (100 µg/ml)* 20 µl8 well sample strips (with frame) 98 well standard control strips* 3* For maximum recovery of the products, centrifuge the original vial after thawing prior to opening the cap.|
|Features & Benefits:||Quick and efficient procedure, which can be finished within 2.5 hours.Innovative fluorometric assay with no need for radioactivity, electrophoresis, and chromatography.Simultaneous quantification of mono-, di-, and tri-methylated H3-K79 with the detection limit as low as 0.4 ng/well and detection range from 5 ng-2 µg/well of histone extracts.The control is conveniently included for quantification of the amount of mono-, di- and tri-methylated H3-K79.Strip microplate format makes the assay flexible: manual or high throughput.Simple, reliable, and consistent assay conditions.|
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