5-mC | 5-methylcystosine (monclonal antibody for MeDIP/IF)
|Data sheet||Product citations||Add review|
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.
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: 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.
Alternative names: 5 Me citidine, 5 Methycytosine, 5 Me Cytidine, methyl CpG.