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>   home   >   Products   >   Primary Antibodies   >   Anti-5-Methylcytidine Antibody, clone 33D3    

Anti-5-Methylcytidine Antibody, clone 33D3

     
  • IF - Anti-5-Methylcytidine Antibody, clone 33D3  ABD11741
    Published customer image:Mouse anti 5-Methylcytidine antibody, clone 33D3 used for the detection of 5-meC in Zea mays root tip cells by immunofluorescence.Image caption:Reduction in 5-meC in TSA- and 5-AC-treated cells. (A) Immunostaining for 5-meC in representative maize root tip cells in interphase or mitosis after TSA or 5-AC treatment. The 'DAPI' panel shows DAPI-stained DNA images, the '5-meC' panel shows immunostained images and the 'Merge' panel shows a combination of blue and green signals. As the untreated control cells progressed into metaphase and early telophase, the amount of 5-meC within mitotic chromosomes increased compared with the amount in interphase cells. The DNA methylation level was reduced in each cell cycle phase after TSA and 5-AC treatments compared with control cells. Bar = 5 µm. (B) Histogram showing mean gray values of the immunostaining signals for 5-meC. The mean gray value of 5-meC signal after treatment with TSA or 5-AC is lower than that of the control. The mean gray values for 5-meC were derived from Additional file 1: Table S3. Error bars represent standard error of the mean.From: Yang F, Zhang L, Li J, Huang J, Wen R, Ma L, Zhou D, Li L. Trichostatin A and 5-azacytidine both cause an increase in global histone H4 acetylation and a decrease in global DNA and H3K9 methylation during mitosis in maize.BMC Plant Biol. 2010 Aug 18;10:178.
  • IF - Anti-5-Methylcytidine Antibody, clone 33D3  ABD11741
    Published customer image:Mouse anti 5-Methylcytidine antibody, clone 33D3 used for the detection of 5-meC in murine embryonic fibroblasts by immunofluorescence.Image caption:The staining pattern of 5meC within nuclei in MEFs. A Phase contrast images of the growth pattern of MEFs in (i) sparse , confluent, and serum deprived confluent cells. Bar = 20 μm. B Typical examples of the 5meC staining patterns of MEFs in (i) proliferative, confluent, and quiescent growth phases. Arrows show cells with a predominantly focal pattern of staining and the other cells show a predominantly diffuse pattern of staining. Scale bar 5 µm. C The proportion (%) of nuclei with predominantly focal 5meC staining in each cell (n, number of nuclei analysed). The focal pattern of 5meC increased with cell proliferation (p
  • IF - Anti-5-Methylcytidine Antibody, clone 33D3  ABD11741
    Published customer image:Mouse anti 5-Methylcytidine antibody, clone 33D3 used for the detection of 5-meC in murine embryonic tissues by immunofluorescence.Image caption:The effect of antigen retrieval procedures on the detection of 5hmC and 5meC in the mouse PN5 stage zygote. Zygotes were collected directly from the oviduct and fixed and prepared for staining. After antigen retrieval steps, zygotes were incubated with primary antibodies for 1 h at room temperature or 18 h at 4°C and each primary antibody was detected by an FITC-labeled secondary antibody. The pronuclei were counter-stained for DNA with PI and the FITC and DNA images merged. Three epitope retrieval approaches were under taken: -H-T, zygotes were not pretreated with either acid or trypsin; +H-T, they were pretreated with acid only; and +H+T, epitope retrieval was by a combination of acid and trypsin treatment. The images are representative of three independent replicates. In each case conditions for staining and imaging were managed so that no signal was detected from the non-immune negative control treatments, and these same conditions used for imaging primary antibody staining. The bar is 5 µm and all images are at the same magnification.From: Li Y, O’Neill C 5'-methylcytosine and 5'-hydroxymethylcytosine Each Provide Epigenetic Information to the Mouse Zygote.PLoS ONE 8(5): e63689.
  • IF - Anti-5-Methylcytidine Antibody, clone 33D3  ABD11741
    Published customer image:Mouse anti 5-Methylcytidine antibody, clone 33D3 used for the detection of 5-meC in murine embryonic fibroblasts by immunofluorescence.Image caption:Staining pattern of 5meC and MBD1 antigens after epitope retrieval acid or acid plus trypsin treatment of confluent MEFs.Fixed and permeabilized MEFs were treated with HCl for 10 or 20A The proportion of nuclei (%) with focal 5meC staining (n, number of nuclei analysed). HCl (p<0.01) and trypsin (p<0.01) treatments revealed an increase in the focal staining of nuclear 5meC. 20 min HCl caused no further increase (p>0.05) after trypsin. B Typical examples of focal and diffuse staining of 5meC staining within the nuclei, yellow and red arrows indicate predominantly diffuse and focal staining, respectively. Non-immune IgG control staining is also shown. C The total level of 5meC staining of treatments shown in A. The results are the mean ± SEM of arbitrary units of optical density increased (**** p<0.0001). D The average number of 5meC foci per cell in A. E The average size (cross-sectional area) of 5meC foci in cells in A. F Representative images of MBD1 staining after 10 min HCl and trypsin 0, 1 or 2 min. Blue and red arrows indicate large and small foci of MBD1, respectively, and yellow arrow shows diffuse staining of MBD1. G The proportion of nuclei with focal MBD1 staining increased after 1 min trypsin (p<0.01). H The total level of MBD1 staining (arbitrary units of optical density) increased after 10 min HCl and 1 min trypsin (p
  • IF - Anti-5-Methylcytidine Antibody, clone 33D3  ABD11741
    Published customer image:Mouse anti 5-Methylcytidine antibody, clone 33D3 used for the detection of 5-meC expression in murine embrryonic tissue by immunofluorescence.Image caption:Pattern of anti-5meC staining in early preimplantation stage embryos. Zygotes were collected directly from the oviduct 16 h – 25 h after the ovulatory injection of hCG. Images show zygotes at PN stage 1-5 (A1-5) and condensing chromosomes , 2-cell , 4-cell and 8-cell . Mitotic zygotes stained with non-immune IgG control is also shown . Antigenic unmasking of 5meC was by brief acid exposure. DNA was counter stained with propidium iodide (PI, Red). Images from these two channels were merged to show co-localization . Images of zygotes were single-equatorial confocal sections (0.77 µm), images of nuclei from 2-cell to 8-cell stage were Z-stacks of multiple sections through each nuclei of the embryo. The images shown here are representative of at least seven independent replicate experiments with at least 10 embryos per observation group per replicated. The scale bars are 10 µm.From: Li Y, O'Neill C Persistence of Cytosine Methylation of DNA following Fertilisation in the Mouse.PLoS ONE 7(1): e30687.
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Product Information
Application
  • Applications Legend:
  • WB=Western Blot
  • IHC=Immunohistochemistry
  • IHC-P=Immunohistochemistry (Paraffin-embedded Sections)
  • IHC-F=Immunohistochemistry (Frozen Sections)
  • IF=Immunofluorescence
  • FC=Flow Cytopmetry
  • IC=Immunochemistry
  • ICC=Immunocytochemistry
  • E=ELISA
  • IP=Immunoprecipitation
  • DB=Dot Blot
  • CHIP=Chromatin Immunoprecipitation
  • FA=Fluorescence Assay
  • IEM=Immunoelectronmicroscopy
  • EIA=Enzyme Immunoassay
WB, IHC-P, IHC-F, IF, FC, E
Host Mouse
Clonality Monoclonal
Isotype IgG1
Clone Names 33D3
Additional Information
Other Species H,Rat,M,B
Shelf Life 18 months from date of despatch.
Target/Specificity Mouse anti-5-Methylcytidine antibody, clone 33D3 recognizes 5-MethylCytidine, a modified base found in the DNA of plants and vertebrates.Methylation of DNA is an epigenetic process that stably alters the expression of genes in cells as they divide and differentiate into specific tissues. The resulting change is normally permanent and unidirectional, preventing a cell from reverting to a stem cell or converting into another type of tissue. In cancer biology, DNA hypermethylation is associated with gene silencing while hypomethylation is reported to be associated with disease progression (Sincic & Herceg, 2011).Mouse anti-5-MeCyd antibody, clone 33D3 is specific to the methylated base and has minimal reactivity to non-methylated cytidine or cytosine (Reynaudet al.1992)Mouse anti-5-MeCyd antibody, clone 33D3 has been reported for use in Methylated DNA immunoprecipitation (MeDIP) (Ponteset al.2007).
Preservative & Stabilisers 0.09% Sodium Azide
Storage Store at +4℃ or at -20 ℃.
PrecautionsAnti-5-Methylcytidine Antibody, clone 33D3 is for research use only and not for use in diagnostic or therapeutic procedures.
Citations (0)

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References

1. Reynaud, C. et al. (1992) Monitoring of urinary excretion of modified nucleosides in cancer patients using a set of six monoclonal antibodies.
Cancer Lett. 61: 255-262. 2. Habib, M. et al. (1999) DNA Global Hypomethylation in EBV - Transformed Interphase Nuclei.
Exp. Cell. Res. 249: 46-53. 3. Hernandez-Blazquez, F. et al. (2000) Evaluation of global DNA hypomethylation in human colon cancer tissues by immunohistochemistry and image analysis.
Gut. 47: 689-693. 4. Giraldo, A. M. et al. (2007) DNA methylation and histone acetylation patterns in cultured bovine fibroblastsfor nuclear transfer.
Mol. Reprod. Dev. 74: 1514-1524. 5. Shen, R. et al. (2009) Reversibility of aberrant global DNA and estrogen receptor-alpha gene methylation distinguishes colorectal precancer from cancer.
Int J Clin Exp Pathol. 2:21-33. 6. Pontes, O. et al. (2007) Postembryonic establishment of megabase-scale gene silencing in nucleolar dominance.
PLoS One. 2: e1157. 7. Yang, F. et al. (2010) Trichostatin A and 5-azacytidine both cause an increase in global histone H4 acetylation and a decrease in global DNA and H3K9 methylation during mitosis in maize.
BMC Plant Biol. 10: 178. 8. Suter, J.D. et al. (2010) Label-free DNA methylation analysis using opto-fluidic ring resonators.
Biosens Bioelectron. 26: 1016-20. 9. Li, Y. and O'Neill, C. (2012) Persistence of cytosine methylation of DNA following fertilisation in the mouse.
PLoS One. 7:e30687. 10. Çelik, S. et al. (2014) The Exit of Mouse Embryonic Fibroblasts from the Cell-Cycle Changes the Nature of Solvent Exposure of the 5'-Methylcytosine Epitope within Chromatin.
PLoS One. 9: e92523. 11. Li, Y. & O'Neill, C. (2013) 5'-Methylcytosine and 5'-hydroxymethylcytosine each provide epigenetic information to the mouse zygote.
PLoS One. 8 (5): e63689. 12. Leter, G. et al. (2014) Exposure to perfluoroalkyl substances and sperm DNA global methylation in Arctic and European populations.
Environ Mol Mutagen. 55 (7): 591-600. 13. Desjobert, C. et al. (2015) Combined analysis of DNA methylation and cell cycle in cancer cells.
Epigenetics. 10 (1): 82-91.1. Sinčić, N & Herceg, Z. (2011) DNA methylation and cancer: ghosts and angels above the genes.
Curr Opin Oncol. 23: 69-76.

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