5-mC | 5-methylcystosine (monclonal antibody for MeDIP/IF)

Product no: AS16 3162
AS16 3162 Clonality: monoclonal  |  Host: Mouse  |  Reactivity:Human, Mouse, other species
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  • Product Info
  • Immunogen: OVA-conjugated molecule: 5-methylcytosine (5-mC)
    Sub class: IgG1
    Host: Mouse
    Clonality: Monoclonal
    Purity: Purified by gel filtration.
    Format: Liquid
    Quantity: 100 g
    Storage: Store at -80C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
    Tested applications: immunofluorescence (IF), MeDIP (methylated DNA Immunoprecipitation (IP)
    Recommended dilution: 05-1 µg (MeDIP), 1: 100 (IF)
  • Reactivity
  • Confirmed reactivity: Human
    Predicted reactivity: Mouse, plants, broad species range
    Not reactive in: No confirmed exceptions from predicted reactivity are currently known
  • Application Examples
  • application information

    MeDIP using monoclonal anti 5-mC (5-methylcystosine) antibodies

    MeDIP was performed using 1 μg fragmented genomic DNA isolated from human blood, the monoclonal antibody against 5-mC and optimized PCR primer sets for qPCR of the indicated regions. 0.2 μg of antibody was used per IP experiment. The graph shows the recovery (expressed as a % of the input DNA, mean of 4 experiments) of the TSH2B gene, known to be methylated and of the promoter of the active GAPDH gene, used as a negative control.

    Methylated DNA immunoprecipitation chart flow'

    Chart flow showing steps of Methylated DNA-immunoprecipiation (MeDIP) method:
    • Prepare genomic DNA from cultured cells
    • Shear genomic DNA
    • Denature the sheared genomic DNA
    • Immunoprecipiate with the antibody against 5-meC
    • Isolate DNA and perform PCR

    immunofluorescence using monoclonal antibodies against 5-mC (5-methylcystosine)

    Immunofluorescence: HeLa cells were stained with the antibody against 5-mC and with DAPI. Cells were fixed with 2.5% formaldehyde for 30’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the 5-mC antibody (left) diluted 1:100 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.
  • Additional Information
  • Additional information: This antibody has been purified by gel filtration, It is supplied in PBS with 0,05 % sodium azide
  • Background
  • Background: 5-mC (5-methylcystosine) is a methylated form of the DNA base cytosine. Methylation may be involved in the regulation of gene transcription.

    Alternative names: 5 Me citidine, 5 Methycytosine, 5 Me Cytidine, methyl CpG.
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