Anti-5-methylcytosine (5-mC) antibody [33D3] (ab10805)


  • Product nameAnti-5-methylcytosine (5-mC) antibody [33D3]
    See all 5-methylcytosine (5-mC) primary antibodies
  • Description
    Mouse monoclonal [33D3] to 5-methylcytosine (5-mC)
  • SpecificityClone 33D3 has been developed to discriminate between the modified base 5-MeCyd and the normal counterpart cytosine.
  • Tested applicationsSuitable for: IHC-P, MeDIP, ChIP, IP, Southern Blot, Dot Blot, Flow Cyt, IHC-Frmore details
    Unsuitable for: ICC
  • Immunogen

    Chemical/ Small Molecule corresponding to 5-methylcytosine (5-mC).

  • General notes

    Storage in frost-free freezers is not recommended. Should this product contain a precipitate microcentrifugation before use.While older lots have performed well in ICC, we have received inconsistent results with the latest lots. Unfortunately, we can no longer guarantee this antibody for use in ICC.




  • FormLiquid
  • Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage bufferpH: 7.4
    Preservative: 0.01% Thimerosal (merthiolate)
    Constituent: 99.99% PBS
  • Concentration information loading...
  • PurityProtein A purified
  • Primary antibody notesAb10805 recognises the modified base 5-methylcytidine found in DNA of plants and vertebrates. DNA methylation is a DNA modification process, which is involved in the control of gene expression. Reports suggest that in tumours, DNA is frequently globally hypomethylated compared to the DNA from normal tissue.
  • ClonalityMonoclonal
  • Clone number33D3
  • MyelomaSp2/0-Ag14
  • IsotypeIgG1
  • Light chain typekappa
  • Research areas


Our Abpromise guarantee covers the use of ab10805 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
MeDIP Use at an assay dependent concentration. PubMed: 17845077Used 5ul in 200ul reaction with 2ug digested genomic DNA from Drosophila.
ChIP Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
Southern Blot 1/200.
Dot Blot Use at an assay dependent concentration. PubMed: 24386123
Flow Cyt Use at an assay dependent concentration.

Use 10ul of working dilution to label 1000000 cells in 100ul. (see Habib, M. et al. (1999)).
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

IHC-Fr Use at an assay dependent concentration.
  • Application notesIs unsuitable for ICC.
  • Target

    • FormThe native context of double-stranded DNA may obstruct antibody binding to 5-methylcytosine. For successful detection of 5-methylcytosine, we recommend that the DNA is denatured to make the nucleotides accessible for the antibody. Denaturing methods vary depending on each application.
    • Alternative names
      • 5 m C antibody
      • 5 mC antibody
      • 5 me C antibody
      • 5 Me Cytidine antibody
      • 5 Me Cytosine antibody
      • 5 meC antibody
      • 5 Methycytosine antibody
      • 5 Methyl Cytidine antibody
      • 5 Methyl Cytosine antibody
      • 5 Methyl-Cytidine antibody
      • 5 Methyl-Cytosine antibody
      • 5 MethylCytidine antibody
      • 5 MethylCytosine antibody
      • 5-mC antibody
      • 5-Me Cytidine antibody
      • 5-Me Cytosine antibody
      • 5-meC antibody
      • 5-Methyl Cytidine antibody
      • 5-Methyl-Cytidine antibody
      • 5-Methyl-Cytosine antibody
      • 5-MethylCytidine antibody
      • 5-MethylCytosine antibody
      • 5mC antibody
      • methyl CpG antibody
      see all

    Anti-5-methylcytosine (5-mC) antibody [33D3] images

    • ab10805 staining 5-Methyl Cytidine in pig embryo (15 to 17 days) tissue section by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue underwent fixation in paraformaldehyde, heat mediated antigen retrieval in Tris-EDTA buffer, permeabilization in Triton X-100 and blocking in 2% BSA for 10 minutes at 25°C. The primary antibody was diluted, 1/100 (PBS + 2% BSA) and incubated with sample for 1 hour at 25°C. An Alexa Fluor® 488 conjugated donkey polyclonal to mouse at 1/250 dilution, was used as secondary.

      See Abreview

    • Native ChIP analysis using unpurified ab10805 binding 5-methylcytosine (5-mC) in nuclear cell lysate derived from mouse embryonic stem cells. Samples were incubated with primary antibody (2µg/µg chromatin) for 16 hours at 4°C in a Glycerol IP buffer. Protein binding was detected using real-time PCR.

      Positive control: Magoh2 in HMT KO cells.

      Negative Control: Magoh2 in WT cells.

    • Dot blot carried out using ab10805 (left blot), showing specificity to 5-methylcytosine. Indicated amounts (20–5 ng) of oligonucleotides containing either 5mC (50%) or 5hmC (20 %) were spotted on positive charged nylon membrane and detected either with 5mC (ab10805) or 5hmC antibody. 

    References for Anti-5-methylcytosine (5-mC) antibody [33D3] (ab10805)

    This product has been referenced in:
    • Hu Y  et al. In Ovo injection of betaine affects hepatic cholesterol metabolism through epigenetic gene regulation in newly hatched chicks. PLoS One 10:e0122643 (2015). WB ; Chicken . Read more (PubMed: 25860502) »
    • Bockor VV  et al. Inhibition of DNA methylation alters chromatin organization, nuclear positioning and activity of 45S rDNA loci in cycling cells of Q. robur. PLoS One 9:e103954 (2014). ICC/IF . Read more (PubMed: 25093501) »

    See all 26 Publications for this product

    Product Wall

    ab10805 specifically binds to single modified cytosine so it should bind to 5-Methylcytosine in RNA as well.

    Application Other
    Sample Zebrafish Tissue lysate - other (5 dpf embryo)

    Kathryn Bambino

    Verified customer

    Submitted Feb 03 2015

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Immunoprecipitation
    Sample Rat Tissue lysate - other (mammary gland)
    Total protein in input 20 µg
    Specification mammary gland
    Treatment maternal high fat
    Immuno-precipitation step Other - Dynabeads

    Abcam user community

    Verified customer

    Submitted Feb 13 2013

    Thank you for contacting us. While we have no tested the specificity for 5-hmC in house, customers have reported that this clone does not recognize 5-hmC. Unfortunately, we do not currently have any data regarding this reactivity.

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application ChIP
    Sample Mouse Cell lysate - nuclear (Mouse ES cells)
    Negative control Magoh2 in WT cells
    Specification Mouse ES cells
    Detection step Real-time PCR
    Type Native ChIP (N-ChIP)
    Positive control Magoh2 in HMT KO cells

    Abcam user community

    Verified customer

    Submitted Dec 03 2012

    Thank you for contacting us.

    There are many anti mouse secondary FITC conjugated antibodies available - they are all fine. Please buy the following antibody;

    Read More

    Thank you for contacting us.

    ab10805 should recognize single 5-methyl cytidine.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Thank you very much for contacting us with your question.

    At this time, there is no discount for the 5-methyl cytidine antibody ab10805. We currently are running a 25% discount promotion for our new product lines (non-primary antibodies) such...

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    Thank you, I am very glad to hear it.
    If you have any problems or questions in the future please do let us know.
    Until then, I wish you all the best with your research.

    Thank you for taking time to provide those protocol details and for contacting us. I am sorry to hear this antibody is not providing satisfactory results.
    The details provided will enable us to investigate this case and will provide us with vital ...

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