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Anti-5-methylcytosine (5-mC) antibody [33D3] ab10805 is a mouse monoclonal antibody that is used in 5-methylcytosine (5-mC) IHC and flow cytometry.

- Antibody clone 33D3 has been tried and trusted by researchers since 2005 and is cited in >170 publications


Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-5-methylcytosine (5-mC) antibody [33D3] (AB10805), expandable thumbnail

Publications

Key facts

Isotype

IgG1

Host species

Mouse

Storage buffer

pH: 7.4
Constituents: PBS, 0.876% Sodium chloride

Form

Liquid

Clonality

Monoclonal

Immunogen

  • Chemical / Small Molecule corresponding to 5-Methylcytosine.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IPIHC-PIHC-FrICCFlow CytSB
Modified Nucleic Acid
Expected
Tested
Expected
Not recommended
Expected
Expected

Expected
Expected

Species

Modified Nucleic Acid

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Modified Nucleic Acid

Dilution info

-

Notes

Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.

Expected
Expected

Species

Modified Nucleic Acid

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Modified Nucleic Acid

Dilution info

-

Notes

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.

Expected
Expected

Species

Modified Nucleic Acid

Dilution info

-

Notes

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.

Expected
Expected

Species

Modified Nucleic Acid

Dilution info

1/200

Notes

-

Associated Products

Select an associated product type

4 products for Alternative Product

Alternative names

Recommended products

Anti-5-methylcytosine (5-mC) antibody [33D3] ab10805 is a mouse monoclonal antibody that is used in 5-methylcytosine (5-mC) IHC and flow cytometry.

- Antibody clone 33D3 has been tried and trusted by researchers since 2005 and is cited in >170 publications

Key facts

Isotype

IgG1

Form

Liquid

Clonality

Monoclonal

Immunogen
  • Chemical / Small Molecule corresponding to 5-Methylcytosine.
Carrier free

Yes

Clone number

33D3

Purification technique

Affinity purification Protein A

Specificity

Modified base 5-methylcytidine found in DNA of plants and vertebrates.

Raised against the modified ribonucleoside. Specific for the presence of a methyl group on carbon 5 of the pyrimidine ring.

Light chain type

kappa

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

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.

Abcam is leading the way to address reproducibility in scientific research with our highly validated recombinant monoclonal and recombinant multiclonal antibodies. Search & select one of Abcam's thousands of recombinant alternatives to eliminate batch-variability and unnecessary animal use.

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Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

5-methylcytosine often referred to as 5-mC 5-methyl cytosine or 5-methylcytidine is a methylated form of the nucleobase cytosine found in the DNA of various organisms. This modification typically occurs at the carbon 5 position of the cytosine ring contributing to the epigenetic regulation of gene expression. While the mass of 5-mC itself is relatively small since it is a modified nucleotide its impact on gene regulation is significant. 5-mC is widely expressed in the genome of higher eukaryotes particularly in CpG dinucleotides where it plays a role in mammalian development and cell differentiation.

Biological function summary

Cytosine methylation affects DNA-protein interactions leading to changes in chromatin structure and the regulation of gene activity. 5-methylcytosine is an important player in the chromatin modification complex where it collaborates with other epigenetic marks to control gene expression profiles. This modified nucleotide helps to stabilize gene silencing maintaining the closed chromatin state which inhibits the transcription machinery's access to specific genomic regions. Such silencing is vital during processes like X-chromosome inactivation and imprinting.

Pathways

Researchers have identified 5-mC within two critical pathways: DNA methylation and demethylation cycle and histone modification pathway. In the DNA methylation pathway 5-mC interacts closely with proteins like DNA methyltransferases (DNMTs) which directly add methyl groups to cytosine bases. In the context of histone modification 5-mC influences the binding of methyl-CpG-binding domain (MBD) proteins which read and interpret methylated DNA marks affecting histone modification and DNA accessibility.

Associated diseases and disorders

The dysregulation of 5-methylcytosine patterns associates prominently with cancer and neurological disorders. Abnormal methylation patterns where 5-mC is either hypermethylated or hypomethylated can lead to the silencing of tumor suppressor genes or activation of oncogenes forming a basis for cancer development. In neurological disorders altered 5-mC levels link to conditions such as Rett syndrome where the dysfunction of the associated protein MeCP2 an important reader of 5-mC contributes to the disease pathology.

Product promise

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1 product image

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-5-methylcytosine (5-mC) antibody [33D3] (ab10805), expandable thumbnail
    This image is courtesy of a customer review submitted by Ms Sara Maj Wätjen Hyldig

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-5-methylcytosine (5-mC) antibody [33D3] (ab10805)

    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.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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