ChIP assays were performed using human osteosarcoma (U-2 OS) cells, the antibody directed against DNMT3B (Cat. no. 49-1028), and optimized PCR primer sets. Each ChIP assay used sheared chromatin from 1 million cells and 0.2 µg of DNMT3B antibody or beads only. The immunoprecipitated DNA was analysed at two loci: the MLH1 promoter, which is a positive locus for DNMT3B, and the CDC6 promoter, a negative locus for DNMT3B. This figure shows the recovery expressed as a percent of the input DNA. Left: Recovery of the MLH1 promoter by the DNMT3B antibody or beads only. Right: Recovery of the CDC6 promoter by the DNMT3B antibody or beads only.
|Tested species reactivity||Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Raised against the mouse DNMT3B protein.|
|Contains||<0.1% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|ChIP assay (ChIP)||0.2 ug|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
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 DNA methyltransferase which is thought to function in de novo methylation, rather than maintenance methylation. The protein localizes primarily to the nucleus and its expression is developmentally regulated. Mutations in this gene cause the immunodeficiency-centromeric instability-facial anomalies (ICF) syndrome. Six alternatively spliced transcript variants have been described. The full length sequences of variants 4 and 5 have not been determined.
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