Anti tubulin antibody

Nitrocellulose-coated coverslips were assembled into flow chambers as described (21 (link)) and used within 24 h of preparation. In experiments with rectangular ridges, one of the coverslips contains the imprinted optical adhesive. The volume of the experimental chambers was ≤20 μL. Solutions were introduced in the chamber in the following sequence: 20 μL of 0.05 mg/mL anti-tubulin antibody (Bio-Rad Laboratories) for 5 min, 50 μL of 2 mg/mL casein for 4 min, 4 × 25 μL of 125 nM microtubules supplemented with 2 mg/mL casein and 20 μM taxol for 4 × 1 min, washed with 100 μL of 2 mg/mL casein, and 50 μL of final solution with kinesin beads, 2 mM ATP, 2 mM MgCl₂, 50 μM DTT, 20 μM taxol, 5 mg/mL glucose, 1500 units/mL of glucose oxidase, and 0.2 units/mL of catalase. The open ends of the chamber were sealed with vacuum grease to prevent evaporation during the experiment.