5-Methylcytosine (5-mC) antibody (mAb) (Clone A1)

Host / Isotype

Mouse / IgG1

Reactivity

Human, Not Species Specific

Applications

DB, IF, MeDIP

CloneNo.

A1

Cat No : 61479,61480

Synonyms

5-methylcytidine, methyl cytidine, 5 MeCyd,5-mC, 5mC, 5mc, 5-Me, 5 MeC, MeDIP, methylated DNA immunoprecipitation, DNA methylation, monoclonal, mouse, mAb, antibody, antibodies, methylcytosine, stem cell, stem cells, sample

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Validation Data Gallery

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  Next-Gen sequencing data generated using 5-Methylcytosine (5-mC) antibody (mAb) correlates well with CpG density. DNA was enriched from 1 ug of denatured, adaptor ligated human PC9 cell DNA using 2 ul of the 5-Methylcytosine (5-mC) antibody (mAb). MeDIP DNA was sequenced using the Illumina platform to generate 26 million sequence tags. Tags were mapped to generate a whole-genome DNA methylation profile. The image above shows that the enriched regions (purple peaks) correlate well with CpG density (green overlay).

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  5-Methylcytosine (5-mC) antibody (mAb) tested by Methyl-DNA Immunoprecipitation (MeDIP). Methylated DNA derived from the promoter of the APC gene was amplified using 5me-dCTP and PCR. 25pg of methylated DNA was spiked into 500 ng of MseI digested human genomic DNA and subjected to the MeDIP procedure using 5μg of 5-Methylcytidine antibody or control mouse IgG. Real time quantitative PCR to amplify the methylated APC promoter DNA was performed on the immunoprecipitated DNA and results plotted as Fold Enrichment over control IgG.

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  5-Methylcytosine (5-mC) antibody (mAb) tested by dot blot analysis. DNA from the Methylated DNA Standard Kit (Catalog No. 55008) were spotted (5 ng per spot) on to a positively charged nylon membrane and blotted with 5-Methylcytosine antibody (2 ug/ml dilution). Lane 1: single-stranded unmethylated DNA. Lane 2: single-stranded DNA containing 5-methylcytosine. Lane 3: single-stranded DNA containing 5-hydroxymethylcytosine. Lane 4: double-stranded unmethylated DNA. Lane 5: double-stranded DNA containing 5-methylcytosine. Lane 6: double-stranded DNA 5-hydroxymethylcytosine.

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Product Information

• Tested Applications: DB, IF, MeDIP

  Applications Validated by Active Motif: MeDIP: 1 - 2 ug per IP MeDIP-Seq: 1 - 2 ug each DB: 0.5 - 2 ug/ml dilution In MeDIP, the DNA must be denatured when using this antibody. For 5-methylcytosine, we also offer AbFlex 5-methylcytosine Recombinant Antibody (rAb). For details, see Catalog No. 91187.

• Tested Reactivity: Human, Not Species Specific
• Host / Isotype: Mouse / IgG1
• Class: Monoclonal
• Type: Antibody
• Modification: Methylated
• Immunogen: This 5-Methylcytosine (5-mC) antibody was raised against 5-Methyl-cytidine conjugated to KLH and recognizes 5-Methylcytosine.
• Full Name: 5-Methylcytosine (5-mC) antibody (mAb) (Clone A1)
• Synonyms: 5-methylcytidine, methyl cytidine, 5 MeCyd,5-mC, 5mC, 5mc, 5-Me, 5 MeC, MeDIP, methylated DNA immunoprecipitation, DNA methylation, monoclonal, mouse, mAb, antibody, antibodies, methylcytosine, stem cell, stem cells, sample
• GenBank accession number: N/A
• RRID: AB_2793653
• Purification Method: Protein A Chromatography
• Buffer: Purified IgG in PBS with 30% glycerol and 0.035% sodium azide. Sodium azide is highly toxic.
• Storage: Some products may be shipped at room temperature. This will not affect their stability or performance. Avoid repeated freeze/thaw cycles by aliquoting items into single-use fractions for storage at -20°C for up to 2 years. Keep all reagents on ice when not in storage.

Background Information

5-Methylcytosine (5-mC) is a modified base that is found in the DNA of plants and vertebrates. DNA methylation is an epigenetic event in which DNA methyltransferases (DNMTs) catalyze the reaction of a methyl group to the fifth carbon of cytosine in a CpG dinucleotide. This modification helps to control gene expression and is also involved in genomic imprinting, while aberrant DNA methylation is often associated with disease. The 5-methylcytidine antibody (Clone A1) has been developed to discriminate between the modified base and its normal cytosine counterpart, allowing for gene promoter methylation analysis.