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Applications: DB (1:400)
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.
Background: 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.
Concentration: 50 µg/28 µl
Cross Reactivity: Does not cross-react with 5-methylcytosine, 5-hydroxymethylcytosine, or 5-carboxylcytosine.
Description: Polyclonal antibody raised in rabbit against 3-methylcytosine (3-mC) conjugated to BSA.
Format: Aqueous buffer solution
Formulation: PBS containing 0.05% azide and 0.05% ProClin 300.
Immunogen: synthetic peptide
Purification: Protein G purified
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.
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
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