Recombinant Human DNMT1 Protein was determined by SDS-PAGE with Coomassie Blue, showing a band at 250-300 kDa.
The 42 base pair of DNA duplex containing three hemimethylation sites (2.5 µM) was methylated with the recombinant DNMT1 (0.18 mg/mL) including 20 µM SAM in reaction buffer (20 mM Tris-HCl [pH 8.0], 50 mM NaCl, 1 mM EDTA, 3 mM MgCl2, 0.1 mg/mL BSA, and 1 mM DTT) at 37 °C for 1 hr. Then methyltransferase assay was performed using MTase-Glo™ Methyltransferase Assay (Promega, V7601). Enzyme 1-4 represent RP01868LQ, Enzyme 5-8 represent recombinant DNMT1 with a 6×His tag at the N-terminus. (Customer feedback data)
DNMT1 (DNA (Cytosine-5-)-Methyltransferase 1) proteins are involved in DNA methylation in which a methyl group is added to a cytosine residue on DNA, commonly at the C5 position of a CpG dinucleotide. Three families of DNMTs have been identified: DNMT1, DNMT2, and DNMT3 (comprised of DNMT3A and DNMT3B).Methylates CpG residues. Preferentially methylates hemimethylated DNA. Associates with DNA replication sites in S phase maintaining the methylation pattern in the newly synthesized strand, that is essential for epigenetic inheritance. Associates with chromatin during G2 and M phases to maintain DNA methylation independently of replication. It is responsible for maintaining methylation patterns established in development. DNA methylation is coordinated with methylation of histones. Mediates transcriptional repression by direct binding to HDAC2. In association with DNMT3B and via the recruitment of CTCFL/BORIS, involved in activation of BAG1 gene expression by modulating dimethylation of promoter histone H3 at H3K4 and H3K9.
