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Journal of Cell Science, Vol 16, 481-497, Copyright © 1974 by Company of Biologists
Submitted on February 5, 1974
Revised on July 10, 1974
1 Biology Department, York University, Downsview, Toronto, Ontario M3J1P3, Canada
2 Department of Zoology, University of Toronto, Toronto, Ontario, M5S 1A1, Canada
A method for isolating sea-urchin zygote mitotic apparatus (MA) is described which is based on the Filner-Behnke method of isolating brain microtubules. MA were isolated in 50% (v/v) glycerol, 10%(v/v) dimethyl sulphoxide, 5 mM MgCl2, 0.1 mM EGTA, and 5 mM Sørensen's phosphate buffer at a final pH of 6.8. MA stored at room temperature in isolation medium had stable birefringence, stable microtubules and stable solubility properties (in 0.5 M KCl) over a period of 10 days to 2 weeks. These MA also seem to have more dry matter per volume than do MA isolated using hexylene glycol. The biggest disadvantages of the method are that zygotes often are difficult to lyse, and that cytoplasmic debris the same size as the MA sometimes contaminates the MA pellet.
Submitted on February 5, 1974
This article has been cited by other articles:
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  M. Stearns, J. Connolly, and D. Brown Cytoplasmic microtubule organizing centers isolated from Polytomella agilis Science, January 16, 1976; 191(4223): 188 - 191. [Abstract] [PDF]
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