Anti-5-methylcytosine (5-mC) antibody [5MC-CD] (ab73938)


  • Product name
    Anti-5-methylcytosine (5-mC) antibody [5MC-CD]
    See all 5-methylcytosine (5-mC) primary antibodies
  • Description
    Mouse monoclonal [5MC-CD] to 5-methylcytosine (5-mC)
  • Host species
  • Specificity
    ab73938 reacts with DNA with 5-Methylcytosine (methylated DNA).
  • Tested applications
    Suitable for: Southern Blot, ICC/IF, Flow Cytmore details
  • Immunogen

    Chemical/ Small Molecule corresponding to 5-methylcytosine (5-mC) conjugated to Bovine Serum Albumin (BSA).

  • Positive control
    • Chlamidomonas me-1 cells.



Our Abpromise guarantee covers the use of ab73938 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
Southern Blot 1/1000.

Immunoblotting - detection of DNA with 5-methylocytosine on nitrocellulose. Use at 1/1000

ICC/IF 1/50 - 1/100. Recommend fixing in 4% PFA followed by denaturation with 70% formamide in 2x SSC and chilling in 70%, 95%, and 99% EtOH for 2 min each. Diluting antibody in SSC (saline sodium citrate) may give optimal results. See protocols from PubMed: 11983892 and 17994007
Flow Cyt 1/100.

ab91545 - Mouse monoclonal IgM, is suitable for use as an isotype control with this antibody.



  • Form
    The 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


  • ab73938 staining 5-Methyl Cytosine in human sperm cells by Flow Cytometry. Cells were fixed with 70% ethanol and permeabilized with 5% tween. The sample was incubated with the primary antibody (1/100 in PBST) for 30 minutes at 27°C. A Alexa Fluor® 488-conjugated goat anti-mouse IgG (1/300) was used as the secondary antibody.


    See Abreview

  • Methylation of chloroplast DNA visualized by immunochemistry. Samples are Chlamidomonas me-1 cells. Left: DAPI-stained cells. Middle: Cells stained with ab73938 and FITC-conjugated 2nd antibody. Right: Merged image. Chloroplast DNA is exclusively methylated in gamete cells
  • ab73938 staining 5-Methyl Cytosine in murine PN5 zygotes by Immunocytochemistry/ Immunofluorescence.

    Cells were fixed with methanol/acetic acid (3:1) and blocked with 30% serum for overnight a 4°C. Samples were incubated with primary antibody (1/100 in diluent) for 18 hours at 4°C before detection with a fluorescent secondary antibody.


This product has been referenced in:
  • Kornicka K  et al. The effects of the DNA methyltranfserases inhibitor 5-Azacitidine on ageing, oxidative stress and DNA methylation of adipose derived stem cells. J Cell Mol Med 21:387-401 (2017). Read more (PubMed: 27998022) »
  • Cantão IH  et al. An Initial Investigation of an Alternative Model to Study rat Primordial Germ Cell Epigenetic Reprogramming. Biol Proced Online 19:9 (2017). Read more (PubMed: 28785173) »

See all 17 Publications for this product

Customer reviews and Q&As

Flow Cytometry
ethanol 70%
Yes - tween 5%
Human Cell (sperm)
Gating Strategy
Cell harvesting/tissue preparation method: percolgradients
Sample buffer: PBS

Ms. Jaleh Barzideh

Verified customer

Submitted Dec 23 2013

Thank you for contacting us.

For the use of ab73938in ICC/IF,I would recommendthe following secondary antibodies:
DyLight 488: ab97007,
DyLight 549: ab97008,
DyLight 594: a...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Immunocytochemistry/ Immunofluorescence
Human Cell (HeLa)
Yes - 0.5% Triton-X100 in PBS

Dr. Kirk Mcmanus

Verified customer

Submitted Feb 07 2012

Thank you for your inquiry. We already removed Southern Blot as application as well as Western Blot. Both applications are somewhat inappropriate since the antibody target is not protein but DNA with methylated cytosine. But I am happy to conf...

Read More

Thanks you for sending the information on you IHC protocol. I am still trying to see if I can find any IHC protocols that are better than others for detected DNA modifications. If I find any I will send them on as soon as I find them. In the meantime, ...

Read More

Thank you for your call today and for letting us know about the trouble with ab73938. As we discussed, please keep me updated of any new results using the antibody at 1:50, and we would appreciate it very much if you send an image of the stainin...

Read More

Thank you for your enquiry. The specificity of the anti-5Methylcytosine antibody has been tested in many published papers included articles by Prof. H. Sano who has done a lot of work on this antibody. Some of them were cited in the references in th...

Read More

Thank you for contacting us and sorry for the delay in getting back to you.

The immunogen used to raise Anti-5-Methyl Cytosine antibody [5MC-CD] (ab73938) is 5-methyl Cytosine conjugated to BSA. It is not 5-methyl Cytosine as part of a ...

Read More


Sign up