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Figure 1. RNA immunoprecipitation using the Diagenode monoclonal antibody directed against psU RNA immunoprecipitation (RIP) was performed on 40 µg total RNA isolated from HeLa cells using 5 µg of the Diagenode monoclonal antibody against psU (Cat. No. C15200247) or with an equal amount of mouse IgG, used as a negative control. The immunoprecipitated RNA was subsequently analysed on a Bioanalyzer. Figure 1A shows the Bioanalyzer profile obtained with the psU antibody (right) The left panel shows the input. Figure 1B shows the gel image for the psU antibody, the IgG negative control and the input (lane 1, 2 and 3, respectively). The marker (in bp) is shown on the left, the position of the 28s and 18s ribosomal RNA is indicated on the right.
Figure 2. RIP using the Diagenode monoclonal antibody directed against psU RIP assays were performed on 40 µg total RNA from human HeLa cells using the Diagenode antibody against psU (Cat. No. C15200247). A titration of the antibody consisting of 1, 2 and 5 µg per RIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control. QRT-PCR was performed with primers for the 18 and 28s rRNA genes, for the RPL19 and ATP5E mRNA, shown to contain a psU residue and for the MB gene, used as a negative control. Figure 2 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).