Diagenode

5-methylcytosine (5-mC) monoclonal antibody cl. b (sample size)

Catalog Number
Format
Price
C15200006-10
10 µg
$90.00
  Bulk order
Other format

Monoclonal antibody raised in mouse against 5-mC (5-methylcytosine) conjugated to ovalbumine.

LotGF-005
Concentration2.1 µg/µl
Species reactivityHuman, mouse, rat, cow, alligator, zebrafish, plants, finch, wide range expected.
TypeMonoclonal
PurityMonoclonal antibody purified by gel filtration.
HostMouse
Storage ConditionsStore at -20°C; for long storage, store at -80°C. Avoid multiple freeze-thaw cycles.
Storage BufferPBS containing 0.05% azide.
PrecautionsThis product is for research use only. Not for use in diagnostic or therapeutic procedures.
Applications Suggested dilution References
MeDIP 0.5 - 1 µg/IP Fig 1
Immunofluorescence 1:1,000 Fig 3

* Please note that of the optimal antibody amount per IP should be determined by the end-user. We recommend testing 0.5-5 µg per IP.

  • Validation data

    MeDIP result

    Figure 1. Methylated DNA immunoprecipitation (MeDIP) results obtained with the Diagenode monoclonal antibody directed against 5-mC
    MeDIP (Methylated DNA immunoprecipitation) was performed on 1 µg fragmented human genomic DNA using 0.2 µg of the Diagenode monoclonal antibody against 5-mC (Cat. No. C15200006) and the MagMeDIP Kit (Cat. No. C02010021). The fragmented DNA was spiked with the internal controls present in the kit (methylated DNA (meDNA) as a positive and unmethylated DNA (unDNA) as a negative control) prior to performing the IP. QPCR was performed with optimized primer sets, included in the kit, specific for the methylated and unmethylated DNA controls, and for a known methylated (TSH2B) and unmethylated (GAPDH) genomic region. Figure 2 shows the recovery expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

    Methylated DNA immunoprecipitation (MeDIP) method

    Figure 2. Methylated DNA immunoprecipitation (MeDIP) method

    • Prepare genomic DNA from cultured cells
    • Shear genomic DNA
    • Denature the sheared genomic DNA
    • Immunoprecipitate with the antibody against 5-meC
    • Isolate DNA and perform PCR

    Immunofluorescence

    Figure 3. Immunofluorescence using the Diagenode monoclonal antibody directed against 5-mC
    HeLa cells were stained with the Diagenode antibody against 5-mC (Cat. No. C15200006) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 1% BSA. The cells were immunofluorescently labelled with the 5-mC antibody (left) diluted 1:1,000 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.

  •  Applications
    Methylated DNA immunoprecipitation
    The Methylated DNA IP (MeDIP) is based on the affinity purification of methylated DNA using an antibody directed against 5-methylcytosine (5-mC) or 5-hydroxymethylcytosine (5-hmC) in the case of hMeDIP. How it works In brief, Methyl DNA IP i... Read more
    IF
    Immunofluorescence: Diagenode offers huge selection of highly sensitive antibodies validated in IF. Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9 HeLa cells transfected with a Cas9 expression vector (... Read more
  •  Documents
    Antibodies you can trust POSTER
    Epigenetic research tools have evolved over time from endpoint PCR to qPCR to the analyses of lar...
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    Datasheet 5mC C15200006 DATASHEET
    Monoclonal antibody raised in mouse against 5-mC(5-methylcytosine) conjugated to ovalbumine.
    Download
    Epigenetic Antibodies Brochure BROCHURE
    More than in any other immuoprecipitation assays, quality antibodies are critical tools in many e...
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  •  Publications

    How to properly cite this product in your work

    Diagenode strongly recommends using this: 5-methylcytosine (5-mC) monoclonal antibody cl. b (sample size) (Diagenode Cat# C15200006-10 Lot# GF-005). Click here to copy to clipboard.

    Using our products in your publication? Let us know!

    TET3 prevents terminal differentiation of adult NSCs by a non-catalytic action at Snrpn.
    Montalbán-Loro R, Lozano-Ureña A, Ito M, Krueger C, Reik W, Ferguson-Smith AC, Ferrón SR
    Ten-eleven-translocation (TET) proteins catalyze DNA hydroxylation, playing an important role in demethylation of DNA in mammals. Remarkably, although hydroxymethylation levels are high in the mouse brain, the potential role of TET proteins in adult neurogenesis is unknown. We show here that a non-catalytic action o...

    Environmental Toxicant Induced Epigenetic Transgenerational Inheritance of Prostate Pathology and Stromal-Epithelial Cell Epigenome and Transcriptome Alterations: Ancestral Origins of Prostate Disease.
    Klukovich R, Nilsson E, Sadler-Riggleman I, Beck D, Xie Y, Yan W, Skinner MK
    Prostate diseases include prostate cancer, which is the second most common male neoplasia, and benign prostatic hyperplasia (BPH), which affects approximately 50% of men. The incidence of prostate disease is increasing, and some of this increase may be attributable to ancestral exposure to environmental toxicants an...

    Genomic integrity of ground-state pluripotency.
    Jafari N, Giehr P, Hesaraki M, Baas R, de Graaf P, Timmers HTM, Walter J, Baharvand H, Totonchi M
    Pluripotent cells appear to be in a transient state during early development. These cells have the capability to transition into embryonic stem cells (ESCs). It has been reported that mouse pluripotent cells cultivated in chemically defined media sustain the ground state of pluripotency. Because the epigenetic patte...

    Developmental origins of transgenerational sperm DNA methylation epimutations following ancestral DDT exposure.
    Ben Maamar M, Nilsson E, Sadler-Riggleman I, Beck D, McCarrey JR, Skinner MK
    Epigenetic alterations in the germline can be triggered by a number of different environmental factors from diet to toxicants. These environmentally induced germline changes can promote the epigenetic transgenerational inheritance of disease and phenotypic variation. In previous studies, the pesticide DDT was shown ...

    Molecular Signatures of Regression of the Canine Transmissible Venereal Tumor.
    Frampton D, Schwenzer H, Marino G, Butcher LM, Pollara G, Kriston-Vizi J, Venturini C, Austin R, de Castro KF, Ketteler R, Chain B, Goldstein RA, Weiss RA, Beck S, Fassati A
    The canine transmissible venereal tumor (CTVT) is a clonally transmissible cancer that regresses spontaneously or after treatment with vincristine, but we know little about the regression mechanisms. We performed global transcriptional, methylation, and functional pathway analyses on serial biopsies of vincristine-t...

    Environmental toxicant induced epigenetic transgenerational inheritance of ovarian pathology and granulosa cell epigenome and transcriptome alterations: ancestral origins of polycystic ovarian syndrome and primary ovarian insufiency.
    Nilsson E, Klukovich R, Sadler-Riggleman I, Beck D, Xie Y, Yan W, Skinner MK
    Two of the most prevalent ovarian diseases affecting women's fertility and health are Primary Ovarian Insufficiency (POI) and Polycystic Ovarian Syndrome (PCOS). Previous studies have shown that exposure to a number of environmental toxicants can promote the epigenetic transgenerational inheritance of ovarian diseas...

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