|Calculated MW||32844 Da|
|Positive Control||HeLa cell nuclear fraction|
|Application & Usage||Western blot analysis (2-4 µg/ml). However, the optimal conditions should be determined individually.|
|Other Names||Methyl-CpG-Binding Domain 2/3|
|Formulation||50 µg in 100 µl PBS containing 0.2% gelatin and 0.05% sodium azide|
|Handling||The antibody solution should be gently mixed before use.|
|Reconstitution & Storage||-20 °C|
|Precautions||MBD2/3 (Clone 106B691) Antibody is for research use only and not for use in diagnostic or therapeutic procedures.|
|Function||Acts as transcriptional repressor and plays a role in gene silencing. Does not bind to DNA by itself (PubMed:12124384). Binds to DNA with a preference for sites containing methylated CpG dinucleotides (in vitro). Binds to a lesser degree DNA containing unmethylated CpG dinucleotides (PubMed:24307175). Recruits histone deacetylases and DNA methyltransferases.|
|Cellular Location||Nucleus. Chromosome. Note=Nuclear, in discrete foci. Detected on chromatin, at promoter regions of active genes|
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Provided below are standard protocols that you may find useful for product applications.
DNA methylation, or the addition of methyl groups to cytosine bases in the dinucleotide CpG, is imperative to proper development and regulates gene expression. The methylation pattern involves the enzymatic processes of methylation and demethylation. A demethylase enzyme has been identified which exhibits demethylase activity associated to a methyl-CpG-binding domain (MBD). The enzyme is able to revert methylated cytosine bases to cytosines within the particular dinucleotide sequence mdCpdG by catalyzing the cleaving of the methyl group as methanol. MeCP2 and MBD1 (PCM1) repress transcription by binding specifically to methylated DNA. MBD2 and MBD4 (also known as MED1) co-localize with foci of heavily methylated satellite DNA and mediate the biological functions of the methylation signal. Surprisingly, MBD3 does not bind methylated DNA either in vivo or in vitro. MBD1, MBD2, MBD3, and MBD4 are expressed in somatic tissues, but the expression of MBD1 and MBD2 is reduced or absent in embryonic stem cells, which are known to be deficient in MeCP1 activity. MBD4 has homology to bacterial base excision repair DNA N-glycosylases/lyases. In some microsatellite unstable tumors, MBD4 is mutated at an exonic polynucleotide tract.
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