ChIP results obtained with the antibody directed against MBD1
ChIP assays were performed using human osteosarcoma (U2OS) cells, the antibody against MBD1 (Cat. # 25303) and optimized PCR primer sets. Sheared chromatin from 1x10⁶ cells and 1.5 µg of antibody were used per ChIP experiment. Beads only were used as a negative IP control. Quantitative PCR was performed with primers for the promoters of the MLH1 gene (used as a positive control) and CDC6 gene (used as a negative control). Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

Determination of the antibody titer
To determine the titer of the antibody, an ELISA was performed using a serial dilution of the antibody directed against human MBD1 (Cat. # 25303), crude serum and flow-through. The plates were coated with the peptide used for immunization of the rabbit. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the purified antibody was estimated to be 1:20,000.

Western blot analysis using the antibody directed against MBD1
Nuclear extracts of HeLa cells (40 µg) were analyzed by Western blot using the antibody against MBD1 (Cat. # 25303) diluted 1:500 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right, the marker (in kDa) is shown on the left. 

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