Recombinant 5mC Antibody [clone RM231] (R20245)

ELISA of single stranded DNA using recombinant 5mC antibody. The plate was coated with streptavidin and then biotinylated single stranded unmethylated DNA, 5-Methylcytosine (5-mC) DNA, and 5-Hydroxymethylcytosine (5-hmC) DNA. A serial dilution of RM231 was used as the primary, and an anti-rabbit IgG AKP secondary.

Direct ELISA of HeLa cell genomic DNA using recombinant 5mC antibody. The plate was directly coated with different concentrations of genomic DNA isolated from HeLa cells. RM231 mAb at 1 ug/ml or 3 ug/ml was used as the primary, and anti-rabbit IgG HRP as secondary.

ICC/IF staining of HeLa cells recombinant 5mC antibody (red). Actin filaments have been labeled with fluorescein phalloidin (green), and nuclei stained with DAPI (blue).

MeDIP was performed using recombinant 5mC antibody 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 1ug 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.

Dot blot of double stranded DNA using recombinant 5mC antibody. 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 RM231 mAb.