Anti-5-Methylcytosine (5mC) Antibody, clone EDL MC-4

32160702

Four types of cytosine modifications have been identified in mammalian DNA, namely 5-methylcytosine (5mC), 5-hydroxymethylcytosine (5hmC), 5-formylcytosine (5fC), and 5-carboxylcytosine (5caC). While 5mC, 5hmC, and 5fC occur in embryonic stem (ES) cells and many cell types in all the major organs (e.g. brain, heart, lung and kidney), 5caC has only been detected in a few cell types at a 10 times lower level than 5fC. The sequential oxidation of 5mC to 5hmC, 5fC, and 5caC catalyzed by the ten-eleven translocation (TET) enzymes has been proposed as a potential route for the demethylation of 5mC back to cytosine (C). Thymine DNA glycosylase (TDG), for example, can excise 5fC and 5caC from DNA as part of the base excision repair (BER) pathway to restore the C-nucleotide. Emerging evidences suggest that the different modifications may be selectively recognized by various proteins, and methyl-CpG-binding protein 2 (MeCP2) binding to 5hmC has been proposed as a means of epigenetic regulation in neurons. The conversion of 5hmC to 5fC, in turn, induces an enhanced binding of DNA-repair-associated proteins to 5fC, which has been proposed as a signal for DNA demethylation.

Target modification is not species-specific
Clone EDL MC-4 targets 5-methylcytosine (5mC), but not 5-hydroxymethylcytosine (5hmC), 5-carboxylcytosine (5caC), or unmodified cytosine (C). A less than 25% cross-reactivity toward 5-formylcytosine (5fC) was observed, which has no impact on 5mC detection by clone EDL MC-4 in cell samples, where 5mC modification level is typically more than 100- to 1,000-fold of 5fC modification level.

BSA-conjugated 5-methylcytosine (5mC).

Dot Blot Analysis: 0.5 µg/mL from a representative lot detected 0.39-100 ng of DNA with 5-methylcytosine (5mC) modification, but not DNA with only 5-hydroxymethylcytosine (5hmC) modification or unmodified cytosine (Courtesy of Jeong-Heon Lee, Ph.D.; Chad Clark; Tamas Ordog, M.D.; Zhiguo Zhang, Ph.D. Epigenomics Development Laboratory, Epigenomics Program, Center for Individualized Medicine, Mayo Clinic, Rochester, Minnesota, USA).

Immunoprecipitation Analysis: 2.5 µg from a representative lot immunoprecipitated 5 ng of DNA with 5-methylcytosine (5mC) modification, but not DNA with only 5-hydroxymethylcytosine (5hmC) modification or unmodified cytosine (Courtesy of Jeong-Heon Lee, Ph.D.; Chad Clark; Tamas Ordog, M.D.; Zhiguo Zhang, Ph.D. Epigenomics Development Laboratory, Epigenomics Program, Center for Individualized Medicine, Mayo Clinic, Rochester, Minnesota, USA).

Research Category
Epigenetics & Nuclear Function
Anti-5-Methylcytosine (5mC), clone EDL MC-4, Cat. No. MABE1081, is a highly specific mouse monoclonal antibody that targets DNA with 5-methylcytosine (5mC) modification and has been tested in Dot Blot and Immunoprecipitation.

Evaluated by Dot Blot analysis of DNA cytosine modifications.

Dot Blot Analysis: 2 µg/mL of this antibody detected 25-100 ng of DNA with 5-methylcytosine (5mC), exhibiting less than 25% cross-reactivity toward 5-formylcytosine (5fC) and no detectable immunoreactivity toward 5-hydroxymethylcytosine (5hmC), 5-carboxylcytosine (5caC), or unmodified cytosine.

Protein G purified.
Purified mouse IgG2a in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Format: Purified

Concentration: Please refer to lot specific datasheet.

100

mouse

purified immunoglobulin

primary antibodies

EDL MC-4, monoclonal

all

dot blot: suitable
immunoprecipitation (IP): suitable

IgG2aλ

ambient

Variable, depending on the size(s) of the DNA molecule(s) with 5-methylcytosine (5mC) modification.

Stable for 1 year at 2-8°C from date of receipt.

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