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

Name: 5-mC | 5-methylcystosine (monclonal antibody for MeDIP/IF)
Brand: IKP
SKU: AS16 3162
Price: 393 EUR
Availability: InStock

AS16 3162 | Clonality: monoclonal | Host: Mouse | Reactivity:Human, Mouse, other species

5-mC | 5-methylcystosine (monclonal antibody for MeDIP/IF) in the group Antibodies Plant/Algal / DNA/RNA/Cell Cycle / Plant Epigenetics/DNA methylation at Agrisera AB (Antibodies for research) (AS16 3162)

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

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 µg

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.

Methylated DNA immunoprecipitation chart flow'

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.

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.

Product citations

immunogen:	OVA-conjugated molecule: 5-methylcytosine (5-mC)
Sub class:	IgG1
Host:	Mouse
Clonality:	Monoclonal
Purity:	Purified by gel filtration.
Format:	Liquid
Quantity:	100 µg
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)
Confirmed reactivity:	Human
predicted reactivity:	Mouse, plants, broad species range
not reactive in:	No confirmed exceptions from predicted reactivity are currently known
Picture (footer):	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.
additional information:	This antibody has been purified by gel filtration, It is supplied in PBS with 0,05 % sodium azide
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.

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