Immunoblotting of DNMT3L polyclonal antibody (Cat # PAB10043) was usedat a 1:1,500 dilution to detect human DNMT3L by western blot after immunoprecipitation using the same antibody. Transfection of U2-OS cells (10cm dish) was accomplished using 3 ug of GAL4-DNMT3L. Protein extraction using IPH at 150 mM. For IP and WB conditions see Fuks et al. (2000). For western blotting the antibody was used at 0.4 ug/mL in TBS milk 2%, BSA 0.5%. Detection occurred using ECL. Note: this antibody works for western blotting only following immunoprecipitation. We have not had good success using the antibody directly for western blotting. The expected molecular weight of human DNMT3L is 43.6 KDa.
CpG methylation is an epigenetic modification that is important for embryonic development, imprinting, and X-chromosome inactivation. Studies in mice have demonstrated that DNA methylation is required for mammalian development. This gene encodes a nuclear protein with similarity to DNA methyltransferases. This protein is not thought to function as a DNA methyltransferase as it does not contain the amino acid residues necessary for methyltransferase activity. However, this protein does stimulate de novo methylation by DNA cytosine methyltransferase 3 alpha and it is thought to be required for the establishment of maternal genomic imprints. This protein also mediates transcriptional repression through interaction with histone deacetylase 1. Alternative splicing results in two transcript variants. An additional splice variant has been described but its biological validity has not been determined. [provided by RefSeq
cytosine-5-methyltransferase 3-like protein,human cytosine-5-methyltransferase 3-like protein