Mono-Methyl Histone H3-K79 Quantification Kit (Colorimetric) is use for measuring mono-methylation of histone H3-K79.
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. H3-K79 di-methylation is a widespread histone modification and is associated with transcriptionally active genes.Increased H3-K79 dimethylation is also found to be involved in some pathological processes such as leukemogenesis in human. The H3-K79 di-methylation can be also changed by inhibiton or activation of HMTs. Thus quantitative detection of di-methyl histone H3-K79 would provide useful information for better understanding epigenetic regulation of gene activation and for developing HMT targeted drugs. The Mono-Methyl Histone H3-K79 Quantification Kit (Colorimetric) provides a tool for measuring di-methylation of histone H3-K79.
For specifically measuring histone H3-K79 mono-methylation using a variety of mammalian cells (human, mouse, etc.) including fresh and frozen tissues, cultured adherent and suspension cells.
For research use only (RUO)
Upon receipt, store C3 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 C1 and C2 contain salt precipitates before using. If so, warm (at room temperature or 37°C) and shake the buffers until the salts are redissolved.
C1 (10X wash buffer) 20 mlC2 (antibody buffer) 12 mlC3 (detection antibody, 1 mg/ml)* 10 μlC4 (color developer) 10 mlC5 (stop solution) 6 mlStandard control (100 μg/ml)* 20 μl8 well sample strips (with frame) 98 well standard control strips*3User guide 1* 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 colorimetric assay with no need for radioactivity, electrophoresis, and chromatography.Specifically capturing mono-methylated H3-K79 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 quantification of the amount of mono-methylated H3-K79.Strip microplate format makes the assay flexible: manual or high throughput.Simple, reliable, and consistent assay conditions.