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

RRID: AB_2793653
Clone: A1
Catalog No: 61479 Format: 100 µg $420 Buy
Catalog No: 61480 Format: 10 µg $105 Buy

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Antibody Type:
Monoclonal
Isotype:
IgG1
Purification:
Protein A Chromatography
Host:
Mouse
Reactivity:
Human, Not Species Specific

Applications

Methyl-DNA Immunoprecipitation Validated Immunofluorescence Validated Dot Blot Validated

Application Notes

Applications Validated by Active Motif:
MeDIP: 1 - 2 µg per IP
MeDIP-Seq: 1 - 2 µg each
DB: 0.5 - 2 µg/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.

Published Applications

The following applications have been published using this antibody. Unless noted above, Active Motif may not have validated the antibody for use in these applications:

IF

View publications that use Active Motif products & services here. Enter the product number(s) to see publications & applications that use this antibody.

Immunogen

This 5-Methylcytosine (5-mC) antibody was raised against 5-Methyl-cytidine conjugated to KLH and recognizes 5-Methylcytosine.

Buffer

Purified IgG in PBS with 30% glycerol and 0.035% sodium azide. Sodium azide is highly toxic.

 

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).

5-Methylcytosine (5-mC) antibody (mAb) tested by Methyl-DNA Immunoprecipitation (MeDIP).
Human DNA (500 ng) digested with MseI was subjected to MeDIP using 2 µg of the 5-Methylcytosine antibody and then analyzed by quantitative real-time PCR with primers specific for ZC3H13 that is normally methylated or Untr1 that is normally unmethylated. The MeDIP'd DNA (enriched DNA) was plotted.

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 µg/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.

Background

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.

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.

Guarantee

This product is guaranteed for 12 months from date of receipt.

This product is for research use only and is not for use in diagnostic procedures.

Application Key

  • ChIP = Chromatin Immunoprecipitation;
  • ChIP = ChIP Sequencing;
  • CUT&RUN = Cleavage Under Targets and Release Using Nuclease;
  • CUT&Tag = Cleavage Under Targets and Tagmentation;
  • DB = Dot Blot;
  • ELISA = Enzyme-linked Immunosorbent Assay;
  • EMSA = Electrophoretic Mobility Shift Assay
  • FC = Flow Cytometry;
  • ICC = Immunocytochemistry;
  • IF = Immunofluorescence;
  • IHC = Immunohistochemistry;
  • IP = Immunoprecipitation;
  • MeDIP = Methyl-DNA Immunoprecipitation;
  • NEU = Neutralizing;
  • WB = Western Blotting