Mouse monoclonal antibodies against α tubulin

Rabbit monoclonal antibody against α-SMA (for histological staining) was from Abcam (Cambridge, UK). Polyclonal rabbit antibody against fibronectin was a generous gift from Dr. Deane Mosher at the University of Wisconsin-Madison. Mouse monoclonal antibodies against α-tubulin, α-SMA and vinculin were from Sigma-Aldrich. Rabbit polyclonal antibody against total fibronectin was from Abcam. Mouse monoclonal antibody against GAPDH was from Santa Cruz Biotech (Santa Cruz, CA). Rabbit polyclonal antibody against total MKL1 was from Bethyl (Montgomery, TX). Mouse monoclonal antibody against E-Cadherin was from BD Biosciences (San Jose, CA).

Doxorubicin, used as a DNA-damaging agent, and 3AB, used as a PARP inhibitor, were purchased from Sigma and Tokyo Chemical Industry Co. (Tokyo, Japan), respectively. AG14361 was a kind gift from Professor Nicola Curtin (Newcastle University, Newcastle Upon Tyne, UK) [22 (link)]. Mouse monoclonal antibodies against KIP1/p27 (BD), mouse monoclonal antibodies against α-tubulin (Sigma, St. Louis, MO, USA), and rabbit polyclonal antibodies against p53 (SANTA CRUZ, Dallas, TX, USA), p21 (SANTA CRUZ), and Skp2 (H-435, SANTA CRUZ) were used for immunoblot analysis. As secondary antibodies, goat anti-mouse IgG HRP conjugate (Nacalai, Kyoto, Japan) and horse anti-rabbit IgG HRP conjugate (Cell Signaling, Danvers, MA, USA) were used.

A rabbit polyclonal antibody from Protein Tech group and a goat polyclonal Ab from Imgenex were used to detect Myo9b. Mouse monoclonal antibodies against α-tubulin and acetylated α-tubulin were purchased from Sigma while mouse monoclonal antibodies against total Src and phospho-Src kinase (Tyr416) were purchased from Upstate/Millipore. An antibody that detects phosphotyrosine residues was from BD Transduction Laboratories. Antibodies to loading controls β-actin and GAPDH were purchased from Abcam. Secondary antibodies for chemiluminescent detection were obtained from Jackson Laboratories. PVDF membrane was purchased from GE Healthcare and SuperSignal West Pico detection reagents were purchased from Thermo Scientific.
Rho and Rac activity were measured with protein pull-down Activation Assay kits from Cytoskeleton Inc. Total Rho/Rac levels were compared to active Rho/Rac and measured using Bio-Rad Quantity One software. Rho Inhibitor I (a cell permeant exoenzyme C3 transferase) was purchased from Cytoskeleton Inc.