Mono-Methyl Histone H3-K4 Quantification Kit (Fluorometric) is use for measuring Mono-methylation of histone H3-K4.
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. SET1, SET7/9, Ash1, ALL-1, MLL, ALR, Trx, and SMYD3 are histone methyltransferases that catalyze methylation of histone H3 at lysine 4 (H3-K4) in mammalian cells. H3-K4 monomethylation is associated with silenced euchromatin regions in the genome and may serve as a epigenetic mark and function in gene repression. The H3-K4 mono-methylation can also be changed by inhibiton or activation of HMTs. Thus quantitative detection of mono-methyl histone H3-K4 would provide useful information for better understanding epigenetic regulation of gene activation/repression and for developing HMT-targeted drugs. The Mono-Methyl Histone H3-K4 Quantification Kit (Fluorometric) provides a tool for measuring Mono-methylation of histone H3-K4.
For specifically measuring histone H3-K4 monomethylation 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 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.
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.Specifically capturing mono-methylated H3-K4 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-methylated H3-K4.Strip microplate format makes the assay flexible: manual or high throughput.Simple, reliable, and consistent assay conditions.