Tri-Methyl Histone H4K20 Quantification Kit (Fluorometric) is used for measuring tri-methylation of histone H4K20.
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 transcriptional repression or activation. HMTs transfer 1-3 methyl groups from S-adenosyl-L-methionine to the lysine and arginine residues of histone proteins. PR-SET7, SET9, SUV4.20h, and ASH1are histone methyltransferases that catalyze methylation of histone H4 at lysine 20 (H4K20) in mammalian cells. Tri-methylation of H4K20 acts as a passive feature or structure determinant for chromatin degradation and release, as well as being an Epigenetic marker of early apoptosis. Tri-methylation of H4K20 is also considered as a common hallmark of human cancer. The H4K20 tri-methylation can be changed by inhibition or activation of HMTs. Therefore, quantitative detection of tri-methyl histone H4K20 would pro-vide useful information for better understanding Epigenetic regulation of tumori-genesis and apoptosis, as well as for developing HMT-targeted drugs. Tri-Methyl Histone H4K20 Quantification Kit (Colorimetric) provides a tool for measuring tri-methylation of histone H4K20.
Tri-Methyl Histone H4K20 Quantification Kit (Fluorometric) is suitable for specifically measuring histone H4K20 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 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) 10 mlF2 (antibody buffer) 6 mlF3 (detecting antibody, 1 mg/ml)* 5 µlF4 (fluoro developer)* 12 µlF5 (fluoro enhancer)* 12 µlF6 (fluoro dilution) 4 mlStandard control (100 µg/ml)* 10 µlSignal report solution* 5 µlSignal enhancer* 120 µl8 well sample strips (with frame) 48 well standard control strips 2User guide 1* For maximum recovery of the products, centrifuge the original vial prior to opening the cap.
Compatible Sample Types:
Features & Benefits:
Quick and efficient procedure, which can be finished within 2.5 hours.Innovative fluorometric assay with no need for radioactivity, electrophoresis, or chromatography.Specifically capture tri-methylated H4K20 with the detection limit as low as 1 ng/well and detection range from 10 ng-2 µg/well of histone extracts.The control is conveniently included for quantification of tri-methylated H4K20.Strip microplate format makes the assay flexible: manual or high throughput.Simple, reliable, and consistent assay conditions.