|Product Overview:||Pan-Methyl Histone H3-K27 Quantification Kit (Fluorometric) is use for measuring mono-, di-and tri-methylation of histone H3-K27.|
|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. G9a and polycomb group enzymes such as EZH2 are histone methyltransferases that catalyze methylation of histone H3 at lysine 27 (H3-K27) in mammalian cells. Mono–methylation, a modification enriched at pericentromeric heterochromatin, was observed to be broadly distributed throughout all euchromatic sites and participates in silencing gene expression. Di– and tri-methylations of H3-K27 are the facultative heterochromatin mark which promotes the recruitment of polycomb group proteins for gene silencing. Increased H3-K27 methylation is found to be involved in some pathological processes such as cancer progress. The patterns of H3-K27 methylation can be also changed by inhibition or activation of HMTs. Thus quantitative detection of methyl histone H3-K27 would provide useful information for better understanding epigenetic regulation of gene activation/ repression and for developing HMT-targeted drugs. The Pan- Methyl Histone H3-K27 Quantification Kit (Colorimetric) provides a tool for measuring mono– di– and tri-methylation of histone H3-K27.|
|Applications:||For simultaneously measuring histone H3-K27 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 3 hours.Innovative fluorometric assay with no need for radioactivity, electrophoresis, and chromatography.Simultaneous quantification of mono-, di-, and tri-methylated H3-K27 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-K27.Strip microplate format makes the assay flexible: manual or high throughput.Simple, reliable, and consistent assay conditions.|
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