Agrisera/5-mC | 5-methylcystosine (monclonal antibody for MeDIP/IF)/AS16 3162/

来自 : 发布时间：2024-05-05

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 Format Quantity 100 µg Reconstitution Storage Store at -80°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 MeDIP (methylated DNA immunoprecipitation), immunofluorescence (IF) Related products AS16 3161 | anti-5-mC | 5-methylcystosine, monoclonal antibodiesAS16 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 known at the moment
Additional information		To be added when available
Selected references		To be added when available, antibody released in February 2016.

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:

• 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.

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