Tri-Methyl Histone H3-K9 Quantification Kit (Fluorometric) is use for measuring tri-methylation of histone H3-K9.
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. ESET, G9a, SUV39-h1, SUV39- h2, SETDB1, Dim-5 and Eu-HMTase are histone methyltransferases that catalyze methylation of histone H3 at lysine 9 (H3-K9) in mammalian cells. H3-K9 trimethylation mediates heterochromatin formation by forming a binding site for HP and is a stable heterochromatin mark which promotes gene silencing. Increased H3-K9 tri-methylation is also found to be involved in some pathological processes such as cancer progression. The H3-K9 tri-methylation can be also changed by inhibiton or activation of HMTs. Thus quantitative detection of tri-methyl histone H3-K9 would provide useful information for better understanding epigenetic regulation of gene activation/silencing and for developing HMT-targeted drugs. The Tri-Methyl Histone H3-K9 Quantification Kit (Colorimetric) provides a tool for measuring tri-methylation of histone H3-K9.
For specifically measuring histone H3-K9 tri-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 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 fluorometic assay with no need for radioactivity, electrophoresis, and chromatography.Specifically capture tri-methyl H3-K9 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 tri-methylated H3-K9.Strip microplate format makes the assay flexible: manual or high throughput.Simple, reliable, and consistent assay conditions.