Fitc coupled anti α tubulin antibody clone dm1a

Indirect immunofluorescence was performed with the anti-EB3 antibody (EPR11421(B), Abcam), anti-acetylated tubulin antibody (Merck millipore), anti-detyrosinated tubulin antibody (Abcam) and anti-mouse antibody Alexa 568 nm (Molecular Probes); and FITC-coupled anti-α-tubulin antibody (clone DM1A; Sigma-Aldrich). Images of cells were captured with Leica DM-IRBE microscope. All images were acquired using Metamorph software (Molecular Devices) at identical acquisition settings, and were processed using Image J software. Mean EB3 comet area was analyzed with Metamorph software on at least 10 representative cells per condition and 3 independent experiments were conducted. Data were expressed as mean ± SEM and statistical analysis was performed using Student’s t-test.

Indirect immunofluorescence was performed as previously described [5 (link)] by using the anti-EB1 antibody and anti-mouse antibody Alexa 568 nm (Molecular Probes); and FITC-coupled anti-α-tubulin antibody (clone DM1A; Sigma-Aldrich). Cells were observed using a Leica DM-IRBE microscope, 100X magnification. All images were acquired using Metamoph software (Molecular Devices, Sunnyvale, CA) at identical acquisition settings, and were processed using Image J software. After cell lysis, 30 μg of total protein were loaded into a 12% SDS-PAGE gel. Anti-EB1 antibody, anti-α-tubulin and anti-mouse IgG-horseradish peroxidase (Jackson Immunoresearch) were used. Chemiluminescence detection kit (Millipore) was used for visualization of protein bands. Chemiluminescent signal was acquired on a G:BOX imaging system (Syngene, Cambridge, UK) and quantification was done with Image J software.