Di-Methyl Histone H3-K4 Quantification Kit (Fluorometric) is use for measuring di-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 di-methylation is associated with transcriptionally permissive euchromatin regions in the genome and may serve as a epigenetic mark in euchromatin and mediates activated transcription. Increased H3-K4 di-methylation is also found to be involved in some pathological processes such as cancer progression. The H3-K4 di-methylation can be also changed by inhibiton or activation of HMTs. Thus quantitative detection of di-methyl histone H3-K4 would provide usefulinformation for better understanding epigenetic regulation of gene activation and for developing HMT-targeted drugs. The Di-Methyl Histone H3-K4 Quantification Kit (Fluorometric) provides a tool for measuring di-methylation of histone H3-K4.
For specifically measuring histone H3-K4 di-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; (2) Store all other components at 4°C away from light. The kit should be stable for 6 months from the shipment date, 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 re-dissolved.
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 dilutionl)* 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 prior to opening the cap.
Features & Benefits:
Quick and efficient procedure, which can be finished within 2.5 hours.Innovative fluorometric assay without need for radioactivity, electrophoresis, and chromatography.Specifically capture di-methylated H3-K4 with the detection limit as low as 0.2 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 di-methylated H3-K4.Strip microplate format makes the assay flexible: manual or high throughput.Simple, reliable, and consistent assay conditions.