Horseradish peroxidase hrp conjugated donkey anti mouse igg

The following antibodies were used for immunofluorescence (IF) analysis and Western blotting (WB): rat monoclonal anti-α-tubulin (1:800 for IF, 1:1000 for WB; MCA78G, Bio-Rad Laboratories, Hercules, CA, USA), rabbit monoclonal anti-phospho-Akt (Ser473) (1:2000 for WB; D9E, Cell Signaling Technology, Danvers, MA, USA), mouse monoclonal anti-γ-tubulin (1:400 for IF; GTU-88, Sigma-Aldrich), and rabbit polyclonal anti-cleaved caspase-3 (1:500 for IF; 1:500 for WB; Asp175, #9661, Cell Signaling Technology). In terms of secondary antibodies, Alexa Fluor 488- or 555-labeled, donkey anti-rabbit or goat anti-rat IgG (1:400–1:800; Life Technologies, Waltham, MA, USA) was used for IF. Horseradish peroxidase (HRP)-conjugated donkey anti-mouse IgG (1:4000, 715-035-151), donkey anti-rabbit IgG (1:4000, 711-035-152), and donkey anti-rat IgG (1:4000, 712-035-153) antibodies were purchased from Jackson ImmunoResearch (West Grove, PA, USA) and were used as secondary antibodies for WB.

Commercial human Topo I siRNA was used in this study (human Top1 siRNA, sc-36694; Santa Cruz Biotechnology). TOP1 was analysed by western blotting using a mouse anti-Topo I antibody (C-21) (1:10,000, sc-32736; Santa Cruz Biotechnology). Commercial human MUS81 siRNA was used in this study (human MUS81 siRNA, L-016143-01-0005; Horizon). MUS81 was analysed by western blotting using a mouse anti-MUS81 antibody (MTA30 2G10/3) (1:10,000, ab-14387; Abcam). As a loading control, α-tubulin was analysed with mouse anti-α-tubulin antibody (10G10) (1:10,000, 017-25031; Fujifilm). Horseradish peroxidase (HRP)-conjugated donkey anti-mouse IgG (1:10,000, 715-035-150; Jackson ImmunoResearch Laboratories) and HRP-conjugated anti-rabbit IgG (1:10,000, 711-035-152; Jackson ImmunoResearch Laboratories) were used as secondary antibodies. The bands were visualised with Chemi-Lumi One Super (Nacalai Tesque) and analysed with LAS4000 Mini (GE Healthcare Life Science).