covalent immobilization of microtubules on glass surfaces for molecular motor force measurements and_0.doc

covalent immobilization of microtubules on glass surfaces for molecular motor force measurements and 导读：就爱阅读网友为您分享以下“covalent immobilization of microtubules on glass surfaces for molecular motor force measurements and_图文”的资讯，希望对您有所帮助，感谢您对92的支持! 2.Add 1 μL of 25 mM Mg-GTP and 2.5 μL DMSO to the tubulin; incubate at 37°C for 15 min. Mix 2 μL Cy3 monofunctional NHS ester with the polymerized tubulin and incubate at 37°C for 40 min in the dark (see Note 38). Transfer the mixture to a clean 0.2 mL TLA-100 tube. Remove the excess dye by centrifugation for 5 min at 37°C at 37,000 rpm (53,000 rcf, average; k-factor48.1). Carefully remove the supernatant, then add 40 μL cold PM buffer to resuspend the pellet. Incubate on ice for 15 min to depolymerize the MTs. Cool rotor to 4 °c. Spin the solution for 5 min at 4 °C at 37,000 rpm (53,000 rcf, average; k-factor 48.1) to remove insoluble fraction. Move supernatant to a clean new tube, add 2 μL of 25 mM Mg-GTP and 5 μL DMSO, and then incubate at 37°C for 15 min in the dark. Warm rotor to 37 °c. Spin the solution for 5 min at 37°C at 37,000 rpm (53,000 rcf, average; k-factor 48.1).