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5-Methylcytosine (5-mC) Monoclonal Antibody [33D3]

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For antibody-based applications requiring high specificity to methylated cytosines

Citations (52) | Write a Review
Methylated DNA successfully captured by 5-methylcytosine antibody (anti-5mc) clone 33D3 during MeDIP. Fig. 2. Immunofluorescence staining with 5-methylcytosine antibody (anti-5mc) clone 33D3.
Synthesized oligos containing different numbers of 5-methylcytosines were captured with the Clone 33D3 anti-5-methylcytosine antibody (Cat No. A-1014) and then colorimetrically detected. The results show that the oligos containing as few as two 5-mCs can still be captured and oligos with four or more 5-mCs can be fully captured by the antibody.
Synthesized oligos containing different numbers of 5-methylcytosines were captured with anti-5-methylcytosine antibodies from various companies and then colorimetrically detected for comparison. Results show Epigentek's 5-mC antibody has the highest sensitivity and specificity in capturing methylated DNA fragments.
Application: DB, ELISA, IF, IHC, IP, MeDIP
Clonality: Monoclonal
Conjugate: Unconjugated
Host: Mouse
Isotype: IgG1
Purification: Affinity Purified
Reactivity: Broad Range, Human, Mouse, Rat
Guarantee: 6 months
Catalog No.SizePriceQty
A-1014-01010 µg $69.00 
A-1014-05050 µg $199.00 
A-1014-100100 µg $359.00 
Availability: Usually Ships In 1-2 Days 
Product Overview

Background
5-methylcytosine (5-mC) is formed when DNA methyltransferase (DNMT) catalyzes the addition of a methyl group onto the 5-carbon of the cytosine ring, an epigenetic process known as DNA methylation. 5-mC is considered the "fifth" DNA base and this 5-methylcytosine mouse monoclonal antibody is ideal for discriminating between the unmodified cytosine base (C) and the methylated cytosine base (5-mC) for DNA methylation studies. DNA methylation, the major epigenetic modification of eukaryotic genomes, plays an essential role in mammalian development. DNA methylation of promoter regions leads to inactivation of gene function. Also, DNA methylation status varies according to tissue type, and region-specific DNA hypermethylation and global DNA hypomethylation have been demonstrated to play an important role in tumorigenesis. 

Description
Mouse monoclonal antibody to 5-methylcytosine (5-mC), clone 33D3, MeDIP/ChIP-grade, used in DNA methylation studies.

Concentration
1 mg/ml

Purification
Protein A

Immunogen
Ovalbumin-conjugated 5-methylcytosine (5-mC)

Isotype
IgG1

Specificity
Modified base 5-methylcytosine (5-mC), a broad range of species.

Formulation
Purified IgG in 10 mM phosphate buffer, 150 mM NaCl, pH 7.4.

Storage
4°C, stable for 6 months from the date of shipment. For long-term storage, aliquot and store at -20°C. Avoid repeated freezing and thawing. Multiple freeze/thaw cycles may result in decreased performance.

Handling Recommendations
For maximum recovery of the products, centrifuge the vial prior to opening the cap.

Alternative Names
5-methylcytidine, anti-5-methylcytidine, anti-5-methylcytosine, anti-5mC, anti-5-mC, anti-5meC, anti-5-meC, 5mC, 5meC, 5-meC, 5'-methyl-2'-deoxycytidine, 5MedCyd

Application & Suggested Dilutions*
Dot Blot: 1:1000-1:2000; Immunohistochemistry: 1:100-1:500; Immunofluorescence: 1:100-1:500; ELISA: 1:1000-1:2000; MeDIP: 0.5-1 µg/reaction

*The end user is responsible for determining optimal working dilutions.


Fig. 1. Methylated DNA successfully captured by 5-methylcytosine antibody (anti-5mc) clone 33D3 during MeDIP.


Fig. 2. Immunofluorescence staining with 5-methylcytosine antibody (anti-5mc) clone 33D3.


Fig. 3. Synthesized oligos containing different numbers of 5-methylcytosines were captured with the Clone 33D3 anti-5-methylcytosine antibody (Cat No. A-1014) and then colorimetrically detected. The results show that the oligos containing as few as two 5-mCs can still be captured and oligos with four or more 5-mCs can be fully captured by the antibody.


Fig. 4. Synthesized oligos containing different numbers of 5-methylcytosines were captured with anti-5-methylcytosine antibodies from various companies and then colorimetrically detected for comparison. Results show Epigentek's 5-mC antibody has the highest sensitivity and specificity in capturing methylated DNA fragments.

User Guide & MSDS

[User Guide]*
*Always use the actual User Guide that shipped with your product. Is the above file locked? You can also request user guides by emailing info@epigentek.com along with your contact information and institution name.

Product Citations

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Savell KE et. al. (July 2016). Extra-coding RNAs regulate neuronal DNA methylation dynamics. Nat Commun. 7:12091.

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Rajabi H et. al. (February 2016). Pronuclear epigenetic modification of protamine deficient human sperm following injection into mouse oocytes. Syst Biol Reprod Med. :1-8.

Heyward FD et. al. (January 2016). Obesity Weighs down Memory through a Mechanism Involving the Neuroepigenetic Dysregulation of Sirt1. J Neurosci. 36(4):1324-35.

Costa G et. al. (January 2016). DNA demethylation caused by 5-Aza-2'-deoxycytidine induces mitotic alterations and aneuploidy. Oncotarget. 7(4):3726-39.

Karpova NN et. al. (January 2016). Protocol for Methylated DNA Immunoprecipitation (MeDIP) Analysis Epigenetic Methods in Neuroscience Research. 105:97-114.

Snyder MD et. al. (December 2015). Suppression of ASH2L alters DNA methylation and histone patterns during bovine embryonic development. Reprod Fertil Dev. 28(2):131.

Golding MC et. al. (July 2015). Histone-lysine N-methyltransferase SETDB1 is required for development of the bovine blastocyst. Theriogenology.

Wu W et. al. (July 2015). A Cell Electrofusion Chip for Somatic Cells Reprogramming. PLoS One. 10(7):e0131966.

Kim SH et. al. (June 2015). Egg-specific expression of protein with DNA methyltransferase activity in the biocarcinogenic liver fluke Clonorchis sinensis. Parasitology. :1-11.

Singh P et. al. (May 2015). Hippocampal chromatin modifying enzymes are pivotal for scopolamine- induced synaptic plasticity gene expression changes and memory impairment. J Neurochem.

Zembrzycki A et. al. (May 2015). Postmitotic regulation of sensory area patterning in the mammalian neocortex by Lhx2. Proc Natl Acad Sci U S A.

Winans B et. al. (May 2015). Linking the Aryl Hydrocarbon Receptor with Altered DNA Methylation Patterns and Developmentally Induced Aberrant Antiviral CD8+ T Cell Responses. J Immunol. 194(9):4446-57.

Fang Y et. al. (March 2015). Effects of the TLR4 transgene on reproductive traits and DNA methylation pattern of oocytes in ewes Front Agr Sci Eng. 1(4):314-20.

Leoni C et. al. (February 2015). Reduced DNA methylation and hydroxymethylation in patients with systemic mastocytosis. Eur J Haematol.

Jia G et. al. (December 2014). Spermatozoa cryopreservation alters pronuclear formation and zygotic DNA demethylation in mice. Theriogenology.

Zovkic IB et. al. (September 2014). Histone H2A.Z subunit exchange controls consolidation of recent and remote memory. Nature.

Nguyen AH et. al. (September 2014). Nanoplasmonic biosensor: Detection and amplification of dual bio-signatures of circulating tumor DNA. Biosens Bioelectron.

Heras S et. al. (June 2014). DNA counterstaining for methylation and hydroxymethylation immunostaining in bovine zygotes. Anal Biochem. 454:14-6.

Wang Y et. al. (April 2014). Hypermethylation of the enolase gene (ENO2) in autism. Eur J Pediatr.

Pierron F et. al. (January 2014). Effect of low-dose cadmium exposure on DNA methylation in the endangered European eel. Environ Sci Technol. 48(1):797-803.

Yu Z et. al. (October 2013). Aldosterone reprograms promoter methylation to regulate αENaC transcription in the collecting duct. Am J Physiol Renal Physiol. 305(7):F1006-13.

Day JJ et. al. (October 2013). DNA methylation regulates associative reward learning. Nat Neurosci. 16(10):1445-52.

Carbone DL et. al. (June 2013). Sex and stress hormone influences on the expression and activity of brain-derived neurotrophic factor. Neuroscience. 239:295-303.

Zhang L et. al. (February 2013). Tet-mediated covalent labelling of 5-methylcytosine for its genome-wide detection and sequencing. Nat Commun. 4:1517.

Sato A et. al. (February 2013). CCAAT/enhancer-binding protein-α suppresses lung tumor development in mice through the p38α MAP kinase pathway. PLoS One. 8(2):e57013.

Zhang X et. al. (January 2013). Promoter hypermethylation of ARID1A gene is responsible for its low mRNA expression in many invasive breast cancers. PLoS One. 8(1):e53931.

Liang Y et. al. (October 2012). DNA methylation pattern in mouse oocytes and their in vitro fertilized early embryos: effect of oocyte vitrification. Zygote. :1-8.

Huang SK et. al. (September 2012). Prostaglandin E₂ increases fibroblast gene-specific and global DNA methylation via increased DNA methyltransferase expression. FASEB J. 26(9):3703-14.

Chen L et. al. (July 2012). Naturally occurring endo-siRNA silences LINE-1 retrotransposons in human cells through DNA methylation. Epigenetics. 7(7):758-71.

Yue MX et. al. (July 2012). Abnormal DNA methylation in oocytes could be associated with a decrease in reproductive potential in old mice. J Assist Reprod Genet. 29(7):643-50.

Yu M et. al. (June 2012). Base-resolution analysis of 5-hydroxymethylcytosine in the mammalian genome. Cell. 149(6):1368-80.

Li Y et. al. (May 2012). Plasticity of DNA methylation in mouse T cell activation and differentiation. BMC Mol Biol. 13:16.

Ge C et. al. (May 2012). A simple colorimetric detection of DNA methylation. Analyst. 137(9):2032-5.

Li Y et. al. (March 2012). IL-2 and GM-CSF are regulated by DNA demethylation during activation of T cells, B cells and macrophages. Biochem Biophys Res Commun. 419(4):748-53.

Barra V et. al. (February 2012). Bypass of cell cycle arrest induced by transient DNMT1 post-transcriptional silencing triggers aneuploidy in human cells. Cell Div. 7(1):2.

Zhang Y et. al. (October 2011). Effects of DNMT1 silencing on malignant phenotype and methylated gene expression in cervical cancer cells. J Exp Clin Cancer Res. 30:98.

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Mandrioli M et. al. (January 2011). Composition and epigenetic markers of heterochromatin in the aphid Aphis nerii (Hemiptera: Aphididae). Cytogenet Genome Res. 133(1):67-77.

Yang Y et. al. (October 2010). Identification of methylated regions with peak search based on Poisson model from massively parallel methylated DNA immunoprecipitation-sequencing data. Electrophoresis. 31(21):3537-44.

Kim MS et. al. (October 2009). DNA demethylation in hormone-induced transcriptional derepression. Nature. 461(7266):1007-12.

Weng YI et. al. (September 2009). Methylated DNA immunoprecipitation and microarray-based analysis: detection of DNA methylation in breast cancer cell lines. Methods Mol Biol. 590:165-76.

Chen C et. al. (June 2009). Aberrant DNA methylation in thymic epithelial tumors. Cancer Invest. 27(5):582-91.

Tsuji Y et. al. (May 2009). The developmental potential of mouse somatic cell nuclear-transferred oocytes treated with trichostatin A and 5-aza-2'-deoxycytidine. Zygote. 17(2):109-15.

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Novikova SI et. al. (April 2008). Maternal cocaine administration in mice alters DNA methylation and gene expression in hippocampal neurons of neonatal and prepubertal offspring. PLoS One. 3(4):e1919.

Mandrioli M et. al. (November 2007). Analysis of heterochromatic epigenetic markers in the holocentric chromosomes of the aphid Acyrthosiphon pisum. Chromosome Res. 15(8):1015-22.

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