5-mC | 5-methylcystosine (monclonal antibody for MeDIP/IF)
AS16 3162 | Clonality: monoclonal | Host: Mouse | Reactivity:Human, Mouse, other species

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Product Information
Immunogen
OVA-conjugated molecule: 5-methylcytosine (5-mC)
Host
Mouse
Clonality
Monoclonal
Subclass/isotype
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 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 examples
application information

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:

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.

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