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5-methylcytosine (5-mC) Antibody for ICC/IF (sample size)

カタログ番号
フォーマット
価格
C15200003-10
10 μg
$95.00
  Bulk order
旧フォーマット



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

Lot003
Concentration1.3 µg/µl
Species reactivityHuman, mouse, drosophila, other (wide range): positive.
TypeMonoclonal
PurityProtein A purified monoclonal antibody.
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 * 1-2 µg per IP
Dot Blotting ** 1:600 Fig 1
Immunofluorescence 1:1,000 Fig 2, 3
FISH 1:200 - 1:1,000 Fig 4, 5

* Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-5 µg per IP.

** Dot blot was only performed to demonstrate the specificity. This antibody is not recommended for dot blot on biological samples

  • Validation Data

    Dot Blot

    Figure 1. Dot blot analysis using the Diagenode monoclonal antibody directed against 5-mC
    To demonstrate the specificity of the Diagenode antibody against 5-mC (Cat. No. C15200003), a Dot blot analysis was performed using the hmC, mC and C controls from the Diagenode “5-hmC, 5-mC & cytosine DNA Standard Pack” (Cat. No. AF-101-0002). One hundred to 4 ng (equivalent of 5 to 0.2 pmol of C-bases) of the controls were spotted on a membrane (Amersham Hybond-N+). The antibody was used at a dilution of 1:600. Figure 1 shows a high specificity of the antibody for the methylated control.

    Immunofluorescence

    Figure 2. Immunofluorescence results obtained with the Diagenode monoclonal antibody directed against 5-mC
    HeLa cells were stained with the Diagenode antibody against 5-mC (Cat. No. C15200003) 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 (middle) diluted 1:1,000 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The left panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.

    A.Immunofluorescence

    B. Immunofluorescence

    Figure 3. Immunofluorescence results obtained with the Diagenode monoclonal antibody directed against 5-mC
    Human osteosarcoma (U2OS) cells were stained with the Diagenode monoclonal antibody against 5-mC (Cat. No. C15200003). Cells were fixed with 2.5% PFA in PBS for 30’, permeabilised with 0.5% Triton X-100 for 1 hour and treated with 2N HCl for 1 hour followed by 2 x 5 minutes with 0.1 M borate buffer to depurinate the DNA. After blocking with PBS containing 0.1% Triton X-100 and 1% BSA, the cells were immunofluorescently labelled with the 5-mC antibody diluted 1:500 in blocking solution, followed by a goat anti-mouse antibody conjugated to Alexa488. Figure 3A: cells were immunofluorescently labelled with the 5-mC antibody after incubation of the antibody with 50 µM mCTP (left) or with DAPI (right). Figure 3C: staining of the cells with the 5-mC antibody after incubation of the antibody with 50 µM hmCTP and with DAPI.

    Fish

    Figure 4. FISH using the Diagenode monoclonal antibody directed against 5-mC
    To detect methylated chromosomal regions, FISH was performed on metaphase chromosomes from HeLa cells using the Diagenode monoclonal antibody against 5-mC (Cat. No. C15200003). The cells were blocked in metaphase by treatment with colcemid (0.1 µg/ml) for 1 - 2 hours, fixed overnight at -20°C with ethanol/glacial acetic acid and treated with 2N HCl for 30’ at room temperature. Subsequently, the cells were blocked with PBS containing 1% BSA and 0.1% Triton X-100 and stained 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 chromosomes with DAPI. A merge of the two stainings is shown on the right.

    Fish

    Figure 5. FISH using the Diagenode monoclonal antibody directed against 5-mC
    To detect methylated chromosomal regions, FISH was performed on metaphase chromosomes from HeLa cells using the Diagenode monoclonal antibody against 5-mC (Cat. No. C15200003). The cells were blocked in metaphase by treatment with colcemid (0.1 µg/ml) for 1 - 2 hours, fixed overnight at -20°C with ethanol/glacial acetic acid and treated with 2N HCl for 30’ at room temperature. Subsequently, the cells were blocked with PBS containing 1% BSA and 0.1% Triton X-100 and stained 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 chromosomes with DAPI. A merge of the two stainings is shown on the right.

  •  実験手法
    DB
    Dot blotting 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
    FISH
    FISH Read more
  •  資料
    Datasheet 5-mC C15200003 DATASHEET
    Monoclonal antibody raised in mouse against 5-mC (5-methylcytosine) conjugated to BSA.
    Download
    Epigenetic Antibodies Brochure BROCHURE
    More than in any other immuoprecipitation assays, quality antibodies are critical tools in many e...
    Download
  •  Safety sheets
    5-mC monoclonal antibody for ICC/IF SDS US en Download
    5-mC monoclonal antibody for ICC/IF SDS GB en Download
    5-mC monoclonal antibody for ICC/IF SDS ES es Download
    5-mC monoclonal antibody for ICC/IF SDS DE de Download
    5-mC monoclonal antibody for ICC/IF SDS JP ja Download
    5-mC monoclonal antibody for ICC/IF SDS BE nl Download
    5-mC monoclonal antibody for ICC/IF SDS BE fr Download
    5-mC monoclonal antibody for ICC/IF SDS FR fr Download
  •  出版物

    How to properly cite this product in your work

    Diagenode strongly recommends using this: 5-methylcytosine (5-mC) Antibody for ICC/IF (sample size) (Diagenode Cat# C15200003-10 Lot# 003). Click here to copy to clipboard.

    Using our products in your publication? Let us know!

    Transcriptome-wide distribution and function of RNA hydroxymethylcytosine
    Delatte B, Wang F, Ngoc LV, Collignon E, Bonvin E, Deplus R, Calonne E, Hassabi B, Putmans P, Awe S, Wetzel C, Kreher J, Soin R, Creppe C, Limbach PA, Gueydan C, Kruys V, Brehm A, Minakhina S, Defrance M, Steward R, Fuks F.
    Hydroxymethylcytosine, well described in DNA, occurs also in RNA. Here, we show that hydroxymethylcytosine preferentially marks polyadenylated RNAs and is deposited by Tet in Drosophila. We map the transcriptome-wide hydroxymethylation landscape, revealing hydroxymethylcytosine in the transcripts of many genes, nota...

    RNA biochemistry. Transcriptome-wide distribution and function of RNA hydroxymethylcytosine.
    Delatte B, Wang F, Ngoc LV, Collignon E, Bonvin E, Deplus R, Calonne E, Hassabi B, Putmans P, Awe S, Wetzel C, Kreher J, Soin R, Creppe C, Limbach PA, Gueydan C, Kruys V, Brehm A, Minakhina S, Defrance M, Steward R, Fuks F
    Hydroxymethylcytosine, well described in DNA, occurs also in RNA. Here, we show that hydroxymethylcytosine preferentially marks polyadenylated RNAs and is deposited by Tet in Drosophila. We map the transcriptome-wide hydroxymethylation landscape, revealing hydroxymethylcytosine in the transcripts of many genes, nota...

イベント

  • ChIP workshop
    Diagenode
    Sep 21-Sep 22, 2021
  • DNA methylation workshop: Master MeDIP assay
    Diagenode
    Sep 30, 2021
  • DNA methylation workshop: Become an expert at RRBS
    Diagenode
    Oct 1-Oct 31, 2021
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