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Figure 1 ChIP results obtained with the Diagenode antibody directed against MBD1 ChIP assays were performed using human osteosarcoma (U2OS) cells, the Diagenode antibody against MBD1 (Cat. No. pAb-078-050) and optimized PCR primer sets. Sheared chromatin from 1x10e6 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).
Figure 2 Determination of the antibody titer To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against human MBD1 (Cat. No. pAb-078-050), 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.
Figure 3 Western blot analysis using the Diagenode antibody directed against MBD1 Nuclear extracts of HeLa cells (40 μg) were analysed by Western blot using the Diagenode antibody against MBD1 (Cat. No. pAb-078-050) 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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H19 lncRNA controls gene expression of the Imprinted Gene Network by recruiting MBD1. Monnier P, Martinet C, Pontis J, Stancheva I, Ait-Si-Ali S, Dandolo L The H19 gene controls the expression of several genes within the Imprinted Gene Network (IGN), involved in growth control of the embryo. However, the underlying mechanisms of this control remain elusive. Here, we identified the methyl-CpG-binding domain protein 1 MBD1 as a physical and functional partner of the H19 ...