Anti-5-methylcytosine (5-mC) antibody [33D3] (ab10805)
Mouse monoclonal 5-methylcytosine (5-mC) antibody [33D3]. Validated in IP, IHC, SB, Flow Cyt, ChIP, Dot, MeDIP. Cited in 67 publication(s). Independently reviewed in 12 review(s).
- Datasheet
- References (82)
- Protocols
Overview
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Product name
Anti-5-methylcytosine (5-mC) antibody [33D3]
See all 5-methylcytosine (5-mC) primary antibodies -
Description
Mouse monoclonal [33D3] to 5-methylcytosine (5-mC) -
Host species
Mouse -
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.
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Tested applications
Suitable for: IHC-P, MeDIP, ChIP, IP, Southern Blot, Dot blot, Flow Cyt, IHC-Frmore details
Unsuitable for: ICC -
Species reactivity
Reacts with: Species independent -
Immunogen
Chemical/ Small Molecule corresponding to 5-methylcytosine (5-mC).
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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.
Properties
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Form
Liquid -
Storage instructions
Shipped 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 buffer
pH: 7.40
Preservative: 0.01% Thimerosal (merthiolate)
Constituent: 99.99% PBS -
Concentration information loading... -
Purity
Protein A purified -
Primary antibody notes
Ab10805 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. -
Clonality
Monoclonal -
Clone number
33D3 -
Myeloma
Sp2/0-Ag14 -
Isotype
IgG1 -
Light chain type
kappa -
Research areas
Associated products
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ChIP Related Products
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Compatible Secondaries
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Isotype control
Applications
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)). |
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| IHC-Fr | Use at an assay dependent concentration. |
Target
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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
see all
Images
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-5-methylcytosine (5-mC) antibody [33D3] (ab10805)](https://www.abcam.com/ps/products/10/ab10805/Images/ab10805-18468-anti-5-methylcytosine-5-mc-antibody-33d3-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-5-methylcytosine (5-mC) antibody [33D3] (ab10805)This image is courtesy of an Abreview submitted by Ms Sara Maj Wätjen Hyldigab10805 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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![ChIP - Anti-5-methylcytosine (5-mC) antibody [33D3] (ab10805)](https://www.abcam.com/ps/products/10/ab10805/Images/ab10805-265452-anti-5-methylcytosine-5-mc-antibody-33d3-chip.jpg)
ChIP - Anti-5-methylcytosine (5-mC) antibody [33D3] (ab10805)This image is courtesy of an anonymous 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.
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![Dot Blot - Anti-5-methylcytosine (5-mC) antibody [33D3] (ab10805)](https://www.abcam.com/ps/products/10/ab10805/Images/ab10805-261699-anti-5-methylcytosine-5-mc-antibody-33d3-dot-blot.jpg)
Dot Blot - Anti-5-methylcytosine (5-mC) antibody [33D3] (ab10805)Image from Nettersheim D et al., PLoS One. 2013;8(12):e82881. Fig S1.; doi: 10.1371/journal.pone.0082881 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/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.
Protocols
References
This product has been referenced in:
- Abukiwan A et al. Dexamethasone-induced inhibition of miR-132 via methylation promotes TGF-ß-driven progression of pancreatic cancer. Int J Oncol 54:53-64 (2019). Read more (PubMed: 30387838) »
- Selli C et al. Molecular changes during extended neoadjuvant letrozole treatment of breast cancer: distinguishing acquired resistance from dormant tumours. Breast Cancer Res 21:2 (2019). Read more (PubMed: 30616553) »
