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

AS16 3162 Clonality: monoclonal  |  Host: Mouse  |  Reactivity:Human, Mouse, other species
5-mC | 5-methylcystosine (monclonal antibody for MeDIP/IF) in the group Plant/Algal Antibodies / DNA/RNA/Cell Cycle / Epigenetics/DNA methylation at Agrisera AB (Antibodies for research) (AS16 3162)


381 €

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product information
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.
Immunogen OVA-conjugated molecule: 5-methylcytosine (5-mC)
Host Mouse
Clonality Monoclonal
Clone IgG1
Purity Purified by gel filtration
Format Liquid
Quantity 100 ĩg
Storage Store at -80°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.
Tested applications immunofluorescence (IF), MeDIP (methylated DNA Immunoprecipitation (IP)
Related products AS16 3161 | anti-5-mC | 5-methylcystosine, monoclonal antibodies

AS16 3163 | anti-5-mC | 5-methylcystosine (monclonal, clone 33D3)

collection of antibodies to epigenetics
Additional information This antibody has been purified by gel filtration. It is supplied in PBS with 0.05 % sodium azide.
application information
Recommended dilution 05-1 µg (MeDIP), 1: 100 (IF)
Expected | apparent MW
Confirmed reactivity Human
Predicted reactivity Mouse, plants, broad species range
Not reactive in No confirmed exceptions from predicted reactivity are currently known.
Additional information
Selected references To be added when available, antibody released in February 2016.

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.

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