Biochemical assay of microtubule mean length
โ Scribed by Fabienne Pirollet; Didier Job; Robert L. Margolis
- Book ID
- 102628686
- Publisher
- Elsevier Science
- Year
- 1987
- Tongue
- English
- Weight
- 492 KB
- Volume
- 167
- Category
- Article
- ISSN
- 0003-2697
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โฆ Synopsis
We present a method for the rapid determination of microtubule mean length in vitro. This method rests on mathematical analysis of the rate of polymer disassembly induced by the introduction of calcium at a known concentration. The rate of disassembly is monitored in our assay by filter trapping of residual microtubule polymers, which contain a radioactive tracer, [3H]GTP. We show that the assay is accurate and reproducible, by comparison with physical measurement of lengths from electron micrographs. Furthermore, we show that the assay can be used to determine rapid shifts in polymer length induced in polymer populations that exhibit "dynamic instability".
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There continues to be intense interest in the microtubule cytoskeleton; the assembly, structure and regulation of microtubules; and the numerous motors and accessory proteins that control cell cycle, dynamics, organization and transport. The field continues to grow and explore new aspects of these i