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 cross-reactivity 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 | ≥ 1:10,000 |
| Immunoblotting | Yes | N/A * |
| Immunofluorescence | Yes | N/A * |
| Immunoprecipitation (IP) of Methylated DNA | Yes | 2-4 µg per IP |
* N/A = Data not available.
Specifications
| Format | Purified |
|---|---|
| Source | Mouse |
| Storage | Store at 0-4°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 0.01% Thimerosal as a preservative. |
| Clone | 10G4 |
| Isotype | IgG1 |
| Product Type | Monoclonal Antibody |
Methylated DNA is efficiently enriched using the 5-Methylcytosine Monoclonal Antibody. DNA was immunoprecipitated using the mouse Anti-5-Methylcytosine 10G4 Antibody from a mixed methylated/non-methylated DNA population. Methylated DNA can be cut with NcoI whereas non-methylated DNA is resistant to NcoI digestion. The DNA (post-IP) was subsequently amplified by PCR and digested with NcoI. Products were then separated in a 2.0% (w/v) agarose/TAE/EtBr gel. The image above demonstrates specific enrichment of methylated versus non-methylated DNA by the Anti-5-Methylcytosine 10G4 Antibody.
