Anti–3–mC polyclonal antibody

Catalog #: 25200
Size: 50 µg
Host Species: Rabbit
Clonality: polyclonal
3–methylcytosine arises by erroneous methylation of DNA through spontaneous exposure to endogenous S–adenosyl methionine (SAM). Since the N3–position in cytosine is protected in double stranded DNA, this phenomenon mainly occurs in single stranded DNA. The resulting 3–mC is toxic and mutagenic and increased levels of 3–mC seem to impair cell proliferation. Therefore, it has to be repaired by the cell. This is established by oxidative demethylation, which is catalyzed by the AlkB protein.
Polyclonal antibody raised in rabbit against 3–methylcytosine (3–mC) conjugated to BSA.
Assay Conditions:

Dot blot analysis using the antibody directed against 3-mC
To demonstrate the specificity of the antibody against 3-mC (cat. No. 25200), a Dot Blot analysis was performed using synthetic oligonucleotides containing different modified bases. 150 and 50 pmol of the respective oligo’s were spotted on the membrane. The antibody was diluted 1:400 in PBS-T containing 10 % skimmed milk and 1% BSA. Figure 1 shows a high specificity of the antibody for the oligonucleotide with the 3-methylcytosine modification.

50 µg/28 µl
PBS containing 0.05% azide and 0.05% ProClin 300.
Species Reactivity:
Human, mouse, broad range
Cross Reactivity:
Does not cross–react with 5–methylcytosine, 5–hydroxymethylcytosine, or 5–carboxylcytosine.
Protein G purified
synthetic peptide
Aqueous buffer solution
Storage / Stability:

Store at –80°C for up to 2 years. Centrifuge after first thaw to maximize product recovery. Aliquot to avoid repeated freeze/thaw cycles. Aliquots may be stored at –20°C for at least one month.

DB (1:400)
The optimal antibody concentration should be determined by the end–user.
Avoid freeze/thaw cycles
Scientific Category:
Product Type:
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