Peroxidase linked anti mouse immunoglobulin g

Rabbit anti UNC-45A (Protein Tech, 1956-1-AP) raised against the C-terminus of the human UNC-45A (Protein Tech, 1956-1-AP) was used for Western blot and immunofluorescence as we have previously described (Habicht et al., 2019a; Mooneyham et al., 2019) . Mouse polyclonal anti UNC-45A raised against full length UNC-45A (Abnova, H00055898-B01P) was used for Western blot to detect recombinant UNC-45A and its mutants (Figure 3B). Mouse anti-αtubulin (Sigma-Aldrich, T6074), for Western blot, rabbit polyclonal α -tubulin (Abcam, ab18251), mouse monoclonal anti-detyrosinated-α-tubulin (EMD Millipore AB3201), rabbit monoclonal anti-GFP (Thermo Fisher Scientific G10362), mouse monoclonal anti-flag (Sigma-Aldrich, F1804), Peroxidase-linked anti-mouse immunoglobulin G and peroxidase-linked antirabbit immunoglobulin G (both Cytiva, formerly known as GE Healthcare Bio-Sciences NA931 and NA934), Alexa Fluor 594-conjugated Donkey Anti-Mouse IgG (1:250) and FITCconjugated Goat Anti-Rabbit IgG (1:200; both Jackson ImmunoResearch Laboratories 715-585-150 and 111-095-003). Paclitaxel was purchased from Teva Pharmaceuticals. Blebbistatin was purchased from Sigma-Aldrich, St. Louis, MO. Tubulin Tracker Deep Red was purchased from Thermo Fisher Scientific (Waltham, MA) cat # T34077.