Alexa fluor 546 igg

All antibodies used for Western blot and immunofluorescence analyses were obtained from commercial sources. Primary antibodies used were anti-E-cadherin (BD Bioscience, NJ, USA), anti-β-catenin (BD Bioscience), anti-active B-catenin (EMD Millipore, Billerica, MA, USA) anti-vimentin (Abcam, Cambridge, MA, USA), anti-fibronectin (Sigma-Aldrich, St. Louis, MO, USA), anti-GFP (Invitrogen, Waltham, MA, USA), anti-β-actin (Sigma-Aldrich), anti-Slug (Cell Signaling Technology, Danvers, MA, USA), anti-FSP-1 (NeoMarkers, Fremont, CA, USA), anti-phosphoAkt1 Ser473 (Cell Signaling Technology), anti-phosphoAkt2 Ser474 (Novus Biologicals Littleton, CO, USA), anti-total Akt1 (Cell Signaling Technology), anti-total Akt2 (Cell Signaling Technology), and rat polyclonal anti-MMP9 (EMD Millipore). β-actin protein levels were used as a control for adequacy of equal protein loading. Western blot membranes were developed using an appropriate Horseradish peroxidase (HRP) conjugated goat anti-mouse or rabbit IgG (Bio-Rad Laboratories, Hercules, CA, USA) or anti-rat antibody (Santa Cruz Biotechnology), and the ECL detection system (EMD Millipore). In the immunofluorescence detection secondary goat anti-mouse Alexa Fluor 488 IgG and goat anti-rabbit Alexa Fluor 546 IgG (Invitrogen) were used.