Different reactivity with monoclonal anti‐tubulin antibodies between native and fixed mitotic microtubules in sea urchin eggs

Mikako T. Oka, Takao Arai, Yukihisa Hamaguchi

研究成果: Article査読

10 被引用数 (Scopus)

抄録

The effect on fixation on the reactivities of mitotic microtubules with monoclonal anti‐tubulin antibodies was investigated by the indirect immunofluorescence procedure. All of the seven antibodies used intensely stained mitotic microtubules in sea urchin eggs lysed and fixed with methanol at −20°C, whereas only two of them stained the stabilized microtubules in the lysed eggs before the fixation. The other five did not stain the mitotic microtubules even after microtubule components other than tubulin were removed by treating the lysed eggs with 0.4 M KCl solution containing taxol. These results exclude the possibility that the fixation affects proteins, which interact with microtubules including microtubule‐associated proteins (MAPs) and interfere with the binding of monoclonal antibodies with tubulin, and strongly suggest that the fixation directly affects the three‐dimensional conformation of tubulin Furthermore, microinjection of these antibodies indicated the results as follows [combining the results reported previously; Oka et al., 1990: Cell Struct. Funct. 15: 373–378]: The antibodies which stained mitotic microtubules stabilized in the lysed eggs induced disassembly of native mitotic microtubules in the living eggs, but those which did not stain the stabilized microtubules did not disassemble the native microtubules. From these results, it is suggested that the monoclonal antibodies which stain microtubules in the eggs lysed but not fixed are useful for microinjection experiments. © 1994 Wiley‐Liss, Inc.

本文言語English
ページ(範囲)241-249
ページ数9
ジャーナルCell Motility and the Cytoskeleton
29
3
DOI
出版ステータスPublished - 1994
外部発表はい

ASJC Scopus subject areas

  • 構造生物学
  • 細胞生物学

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