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Figure 1. RNA immunoprecipitation using the Diagenode monoclonal antibody directed against Inosine RNA immunoprecipitation (RIP) was performed on 40 µg total RNA isolated from HeLa cells using 1 µg of the Diagenode monoclonal antibody against Inosine (cat. No. C15200251) 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 Inosine antibody (right) The left panel shows the input. Figure 1B shows the gel image for the Inosine 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 Inosine RIP assays were performed on 40 µg total RNA from human HeLa cells using the Diagenode antibody against Inosine (cat. No. C15200251). A titration of the antibody consisting of 1, 2, 5 and 10 µg per RIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control. QRT-PCR was performed with primers for the 18s and 28s rRNA genes. Figure 2 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).
Figure 3. RIP using the Diagenode monoclonal antibody directed against Inosine RIP was performed with 1 µg of the Diagenode antibody against Inosine (cat. No. C15200251) on 40 µg total RNA from human HeLa cells was spiked with an in vitro produced RNA molecule containing Inosine nucleotides as well as an unmodified control RNA (100 ng each). IgG (1 µg/IP) as well as an m6A antibody (C15410208) were used as negative control. QRT-PCR was performed with primers specific for the Inosine and unmodified RNA molecules. Figure 3 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).
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