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5-Methylcytosine (5-mC) antibody (rAb) (Clone RM231)

Host / Isotype

Rabbit / IgG

Reactivity

Human, Not Species Specific

Applications

Dot Blot, ICC, MeDIP

CloneNo.

RM231

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Cat No : 91311 91311

Synonyms

5-methylcytidine, methyl cytidine, 5 MeCyd, 5-Me, 5 MeC, MeDIP, methylated DNA immunoprecipitation, DNA methylation

Validation Data Gallery

  • Dot blot of 5-Methylcytosine (5-mC) antibody (rAb).Dot blot of double stranded DNA using 5-Methylcytosine (5-mC) antibody (RM231). The membrane was pre-spotted with 50, 5, and 0.5 ng/dot of double stranded 5-Hydroxymethylcytosine (5-hmC) DNA, 5-Methylcytosine (5-mC) DNA, and unmethylated DNA. The pre-spotted membrane was then blotted with 5-Methylcytosine (5-mC) antibody at 0.5 ug/ml.

    Dot blot of 5-Methylcytosine (5-mC) antibody (rAb).Dot blot of double stranded DNA using 5-Methylcytosine (5-mC) antibody (RM231). The membrane was pre-spotted with 50, 5, and 0.5 ng/dot of double stranded 5-Hydroxymethylcytosine (5-hmC) DNA, 5-Methylcytosine (5-mC) DNA, and unmethylated DNA. The pre-spotted membrane was then blotted with 5-Methylcytosine (5-mC) antibody at 0.5 ug/ml.

  • MeDIP of 5-Methylcytosine (5-mC) antibody (rAb).MeDIP was performed using 5-Methylcytosine (5-mC) antibody (RM231) at a 2:1 DNA:Ab ratio. 1 ng of unmethylated, 5-Methylcytosine (5-mC) or 5-Hydroxymethylcytosine (5-hmC) DNA standard (897 bp) was spiked in 1 ug of genomic DNA isolated from HeLa cells as the control. Realtime PCR was then performed to determine the capture of DNA standard as in % of input.

    MeDIP of 5-Methylcytosine (5-mC) antibody (rAb).MeDIP was performed using 5-Methylcytosine (5-mC) antibody (RM231) at a 2:1 DNA:Ab ratio. 1 ng of unmethylated, 5-Methylcytosine (5-mC) or 5-Hydroxymethylcytosine (5-hmC) DNA standard (897 bp) was spiked in 1 ug of genomic DNA isolated from HeLa cells as the control. Realtime PCR was then performed to determine the capture of DNA standard as in % of input.

  • Immunofluorescence stain of 5-Methylcytosine (5-mC) antibody (rAb).HeLa cells were stained using 5-Methylcytosine (5-mC) antibody (RM231) antibody at 1 ug/ml (red). Actin filaments have been labeled with fluorescein phalloidin (green), and nuclei stained with DAPI (blue).

    Immunofluorescence stain of 5-Methylcytosine (5-mC) antibody (rAb).HeLa cells were stained using 5-Methylcytosine (5-mC) antibody (RM231) antibody at 1 ug/ml (red). Actin filaments have been labeled with fluorescein phalloidin (green), and nuclei stained with DAPI (blue).

Product Information

Tested Applications Dot Blot, ICC, MeDIP

Validated Applications: DB: 0.5 - 2 ug/ml ICC: 0.5 - 2 ug/ml MeDIP: 0.2 - 2 ug/ml
Tested Reactivity Human, Not Species Specific
Host / Isotype Rabbit / IgG
Class Recombinant
Type Antibody
Immunogen BSA-conjugated 5-methylcytosine.
Full Name 5-Methylcytosine (5-mC) antibody (rAb) (Clone RM231)
Synonyms 5-methylcytidine, methyl cytidine, 5 MeCyd, 5-Me, 5 MeC, MeDIP, methylated DNA immunoprecipitation, DNA methylation
Molecular weight
GenBank accession numberN/A
Purification Method Protein A Chromatography
Buffer PBS with 50% glycerol, 1% BSA, and 0.09% sodium azide. Sodium azide is highly toxic.
Storage Some products may be shipped at room temperature. This will not affect their stability or performance. Avoid repeated freeze/thaw cycles by aliquoting items into single-use fractions for storage at -20°C for up to 2 years. Keep all reagents on ice when not in storage.

Background Information

5-Methylcytosine (5-Methylcytidine) is a modified base that is found in the DNA of plants and vertebrates. DNA methylation is an epigenetic event in which DNA methyltransferases (DNMTs) catalyze the reaction of a methyl group to the fifth carbon of cytosine in a CpG dinucleotide. This modification helps to control gene expression and is also involved in genomic imprinting, while aberrant DNA methylation is often associated with disease.