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- W165089717 abstract "This chapter describes a protocol for the isolation of large quantities of centrosomes from early Drosophila embryos and two protocols for assaying the microtubule-nucleating activity of the isolated centrosomes. These centrosomes display a robust microtubule nucleation activity in vitro, each one generating an array of hundreds of microtubules. These centrosomes have a large amount of pericentriolar material compared to isolated mammalian centrosomes. Other advantages of using Drosophila embryos as a source of centrosomes include the ease of obtaining multigram quantities of embryos in a few hours, the potential for isolating centrosomes from mutant fly stocks, the ability to employ genetics and the unique cell biological properties of live embryos in studying any new proteins identified, and the low cost of maintaining large populations of flies. The yield of active centrosomes from this preparation is typically 1–5 X 107 per sucrose gradient, from the equivalent of 2 g of embryos. Approximately, 5 μg of protein is found in the peak fraction (0.5 ml). Assuming that there is an average of 2500 centrosomes in a Drosophila embryo, and that an embryo is 9.5 μg, the yield is estimated to be an approximately 10%." @default.
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- W165089717 date "1998-01-01" @default.
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- W165089717 title "Chapter 1 Isolation of Centrosomes from Drosophila Embryos" @default.
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- W165089717 doi "https://doi.org/10.1016/s0091-679x(08)61972-x" @default.
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