5-mC | 5-methylcystosine (monclonal antibody)

356 €
AS16 3161 clonality: monoclonal  |  host: mouse  |  reactivity:human, mouse, other species


1 st
Item No:
AS16 3161

Info: Product suggestions Add review
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 BSA-conjugated molecule: 5-methylcytosine (5-mC)
Host Mouse
Clonality Monoclonal
Clone IgG1
Quantity 50 ĩg
Storage Store at -20°C; 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 material adhering to the cap or sides of the tubes.
Tested applications dot blot (Dot), immunofluorescene (IF), immunoprecipiation (IP), FISH (FISH), MeDIP (methylated DNA immunoprecipitation)
Related products AS16 3162 | 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 using Protein A and is present in PBS.
application information
Recommended dilution 1: 600 (Dot), 1-5 µg (IP), 1: 500 (IF), 1: 200-1: 1000 (FISH)
Expected | apparent MW
Confirmed reactivity


Predicted reactivity mouse, broad species range
Not reactive in No confirmed exceptions from predicted reactivity known at the moment
Additional information To be added when available
Selected references To be added when available, antibody released in February 2016.

application example

Dot blot using the monoclonal anti-5-mC (5-methylcystosine) antibodies'
Dot blot: to demonstrate the specificity of the monoclonal antibody against 5-mC, a Dot blot analysis was performed using the hmC, mC and C controls: “5-hmC, 5-mC & cytosine DNA. One hundred to 4 ng (equivalent of 5 to 0.2 pmol of C-bases) of the controls were spotted on a membrane (Amersham Hybond-N+). The antibody was used at a dilution of 1:600.

immunofluorescence with monoclonal antibodies to 5-mC (5-methylcystosine)

Immunofluorescence: human osteosarcoma (U2OS) cells were stained with the monoclonal antibody against 5-mC. Cells were fixed with 2.5% PFA in PBS for 30’, permeabilised with 0.5% Triton X-100 for 1 hour and treated with 2N HCl for 1 hour followed by 2 x 5 minutes with 0.1 M borate buffer to depurinate the DNA. After blocking with PBS containing 0.1% Triton X-100 and 1% BSA, the cells were immunofluorescently labelled with the 5-mC antibody diluted 1:500 in blocking solution, followed by a goat anti-mouse antibody conjugated to Alexa488. Panel (A): cells were immunofluorescently labelled with the 5-mC antibody (left) or with DAPI (right). Panel (B) and (C): staining of the cells with the 5-mC antibody after incubation of the antibody with 50 μM mCTP or hmCTP, respectively.

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

FISH: to detect methylated chromosomal regions, FISH was performed on metaphase chromosomes from HeLa cells using the monoclonal antibody against 5-mC. The cells were blocked in metaphase by treatment with colcemid (0.1 μg/ml) for 1 - 2 hours, fixed overnight at -20°C with ethanol/glacial acetic acid and treated with 2N HCl for 30’ at room temperature. Subsequently, the cells were blocked with PBS containing 1% BSA and 0.1% Triton X-100 and stained with the 5-mC antibody (left) diluted 1:1,000 in blocking solution, followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the chromosomes with DAPI. A merge of the two staining is shown to the right.