| 4 mg/mL Rhodamine-tubulin |
| 100 mM MgCl2 |
| 25 mM GTP |
| DMSO |
| BRB80 = | 80 mM PIPES pH 6.85 1 mM MgCl2 1 mM EGTA |
| BRB80CT = | BRB80 0.2 mg/mL casein 10 micromolar Taxol* *Add Taxol (1 mM in DMSO) while vortexing solution |
| 1. | Mix 10 microliters rhodamine-tubulin, 0.5 microliter MgCl2, 0.5 microliter GTP, 0.6 microliter DMSO, and 0.9 microliter BRB80. The total volume is 12.5 microliters. |
| 2. | Grow for 30 min at 37°C. |
| 3. | Dilute 100-fold into room-temperature BRB80CT. Label these MT100. |
| 1. | These microtubules are 0.32 micromolar and are long: 5 to 20 microns. |
| 2. | Final concentrations:
MgCl2 = 4 mM GTP = 1 mM DMSO = 4.8% |
Contributed by William Hancock, J. Howard Laboratory
If you use methods from the Kinesin site, we ask that you cite the Kinesin Home Page and authors, or the appropriate source publication in your work.
Copyright 1996-2003. All rights reserved.
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Created 11 April 1999 21:10 GMT
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Modified 23 July 2001 19:05 GMT