DNA methylation is a type of chemical modification of DNA that can be inherited and subsequently removed without changing the original DNA sequence. Therefore it is part of the epigenetic code and is also the most well characterized epigenetic mechanism. DNA methylation results in addition of a methyl group to DNA — for example, to the number 5 carbon of the cytosine pyrimidine ring — which involves reduction in gene expression. In adult somatic tissues, DNA methylation typically occurs in a CpG dinucleotide context; non-CpG methylation is prevalent in embryonic stem cells.
Immunocytochemistry (Figure below and Ref.1 & 2) (~50-100 fold dilution)
Immuno-blot detection of DNA with 5-methylocytosine on nitrocellulose (Ref. 3 & 4) (~1000 fold dilution)
Immunogen: 5-Methylcytosine conjugated to bovine serum albumin (Ref 3)
Reactivity: DNA with 5-Methylcytosine (methylated DNA), any species
Form: Purified biotinylated mouse IgM, 1 mg/ml in PBS with 50% glycerol, filter- sterilized. Azide and carrier free
Storage: Ship 4°C, upon arrival aliquot and store at -20°C
Fig.1 Methylation of chloroplast DNA visualized by immunochemistry. Samples are Chlamidomonas me-1 cells. Left: DAPI-stained cells; Middle: Cells stained with anti-5MeC antibody and FITC-conjugated 2nd antibody; Right: Merged image. Chloroplast DNA is exclusively methylated in gamete cells.
Fig.2 Detection of DNA methylation in mouse embryonic stem cells by immunofluorescence staining with the anti-5MeC antibody. Intense 5-methylcytosine staining at pericentromeric regions was seen in the mitotic chromosome and interphase nuclei of ESCs (For details, see Reference 1).
This product has been used in following references and many other publications.
Sharif J et al “The SRA protein Np95 mediates epigenetic inheritance by recruiting Dmnt1 to methylated DNA” Nature 450: 908-912 (2007) PMID: 17994007
Nishiyama R et al “A chloroplast-resident DNA methyltransferase is responsible for hypermethylation of chloroplast genes in Chlamydomonas maternal gametes” PNAS 99: 5925-5930 (2002) PMID: 11983892
Sano H et al “Detection of heavy methylation in human repetitive DNA subsets by a monoclonal antibody against 5-methylcytosine” Biochim Biophys Acta 951:157-65 (1988) PMID: 2847796
Sano H et al “Identification of 5-methycytosine in DNA fragment immobilized on nitrocellulose paper “PNAS 77:3581-3585 (1980) PMID: 6251470