Figure 1. ChIP results obtained with the Diagenode antibody directed against TIP5 ChIP assays were performed using the Diagenode antibody against TIP5 (cat. No. CS-090-100). Chromatin from HEK293T and NIH3T3 cells was formaldehyde cross-linked and sheared with the Bioruptor (Diagenode) to yield fragments with an average length of 200 to 400 bp. ChIP was performed overnight at 4°C with 100 μg sheared chromatin and either 5 μl of the TIP5 antibody or 5 μl IgG which was used as negative IP control. The IP’d DNA was analysed by qPCR with primer sets for the promoter and the coding region of the 28s ribosomal RNA gene. Figure 1 shows the recovery by the TIP5 antibody and by IgG (set to 1), normalised to the input DNA. These results show that, both in HEK293T and in NIH3T3 cells, TIP5 is associated with the promoter, but not with the coding region of the 28srRNA gene.
Figure 2. Western blot analysis using the Diagenode antibody directed against TIP5 Western blot was performed on 150 μg nuclear extract from either NIH3T3 or HeLa cells with the Diagenode antibody against TIP5 (cat. No. CS-090-100), diluted 1:1,000 in PBS containing 5% milk powder and 0.1% Tween- 20. The molecular weight marker is shown on the left, the location of the protein of interest is indicated on the right.