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Product Size Catalog # Price Qty
Anti-5-Methlycytosine 15 ul @ 1 ug/ul (Clone 10G4) 15 µg A3001-15
Anti-5-Methlycytosine 30 ul @ 1 ug/ul (Clone 10G4) 30 µg A3001-30
Anti-5-Methlycytosine 50 ul @ 1 ug/ul (Clone 10G4) 50 µg A3001-50
Anti-5-Methlycytosine 200 ul @ 1 ug/ul (Clone 10G4) 200 µg A3001-200

About Anti-5-Methylcytosine

The mouse Anti-5-Methylcytosine monoclonal antibody (clone 10G4) has been developed to facilitate differentiation between methylated and non-methylated cytosines in DNA. Specificity of this clone is to 5-methylcytosines in single-stranded DNA with no detectable crossreactivity to non-methylated cytosines. The antibody has proven to be a valuable tool in the characterization of DNA methylation and has been successfully used for immunopreciptiation-based assays such as Methylated DNA Immunoprecipitation (MeDIP).

Application Recommended Dilution
ELISA Yes ≥ 1:4,000
Flow Cytometry N/D * N/A *
Immunoblotting Yes ≥ 1:5,000
Immunofluorescence Yes N/A *
Immunohistology N/D * N/A *
Immunoprecipitation (IP) Yes 2-4 µg per IP
Western Blotting N/D * N/A *

* N/A = Data not available. N/D = Not Determined

Format Purified
Source Mouse
Storage Store at 0 to 4° C with prolonged storage at -80° C. Avoid repeated freeze/thawing.
Specificity 5-Methylcytosine in single stranded DNA from any source including human, rat, mouse, plants, etc. and 5-Methylcytidine
Approximate Protein Concentration 1 mg/ml
Buffer Solution Antibody is provided in PBS (Phosphate Buffered Saline, pH 7.4) with 0.05% Sodium azide as a preservative.
Clone 10G4
Isotype IgG1
Product Type Monoclonal Antibody

Methylated DNA is efficiently enriched using the 5-Methylcytosine antibody DNA was immunoprecipitated using the mouse Anti-5-Methylcytosine 10G4 antibody from a mixed methylated/non-methylated DNA population. This population is comprised of a mixture of non-methylated plasmid DNA and a methylated version of the same plasmid containing a point mutation that introduces an NcoI restriction site. After immunoprecipitation of the mixture, the region of DNA containing the restriction site was amplified by PCR, digested with NcoI, and visualized in a 2.0% (w/v) agarose/TAE/EtBr gel. Non-methylated DNA remains un-cut, whereas the methylated DNA is cut by NcoI. The image above demonstrates specific enrichment of methylated versus non-methylated DNA by the Anti-5-Methylcytosine 10G4 antibody.
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Featured Citations

Anti-5mC antibodies were used in a southwestern dot blot assay to detect 5-methylcytosine in genomic DNA from E. coli expressing DNA methyltransferases (MTases) from various bacterial strains. Dot blots were then visualized and the hybridization signals were quantified to compare the DNA methylation activity of the different MTases.

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