CRISPR/Cas9 C-terminal monoclonal antibody

Figure 1. Western blot analysis using the Diagenode monoclonal antibody directed against CRISPR/Cas9
Western blot was performed on protein extracts from HeLa cells transfected with Cas9 using the Diagenode antibody against CRISPR/Cas9 (cat. No. C15200223). The antibody was used at different dilutions. The marker is shown on the left, position of the Cas9 protein is indicated on the right.

Figure 2. Western blot analysis using the Diagenode monoclonal antibody directed against CRISPR/Cas9
Western blot was performed on 20 µg protein extracts from Cas9 expressing HeLa cells (lane 1) and on negative control HeLa cells (lane 2) with the Diagenode antibody against Cas9 (cat. No. C15200223). The antibody was diluted 1:4,000. The marker is shown on the left, position of the Cas9 protein is indicated on the right.

Figure 3. IP using the Diagenode monoclonal antibody directed against Cas9
IP was performed on whole cell extracts (250 & micro;g) from HeLa cells transfected with a Cas9 expression vector (lane 1 and 3), or untransfected cells (lane 2 and 4) using 13 µg of the Diagenode antibody against Cas9 (cat. No. C15200223). The immunoprecipitated proteins were subsequently analysed by Western blot. Lane 3 and 4 show the result of the IP, the input (15 µg) is shown in lane 1 and 2.

Figure 4. Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9
HeLa cells expressing Cas9 under the control of the tight TRE promoter were fixed in methanol at -20°C, permeabilized with acetone at -20°C and blocked with PBS containing 2% BSA. The cells were stained with the Cas9 C-terminal antibody (cat. No. C15200223) diluted 1:200, followed by incubation with a donkey anti-mouse secondary antibody coupled to AF488. Nuclei were counter-stained with Hoechst 33342. Figure 4 shows the result in the presence (left) or absence (right) of doxycycline.