시간 경과 현미경과 분해 굴지 모니터링

Biology

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Summary

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Kueh, H. Y. Monitoring Actin Disassembly with Time-lapse Microscopy. J. Vis. Exp. (1), e66, doi:10.3791/66 (2006).

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Abstract

Protocol

  1. 로 비디오에 나타난 흐름 챔버를 구성. parafilm 밀봉이 꽉 있는지 확인하고 세탁 coverslips을 사용합니다.
  2. 50-10 '에 대한 챔버에 품어 굴지 바인딩 요원.
  3. 차단 단백질과 유리 coverslip에 비 특정 바인딩 사이트를 차단합니다.
  4. 버퍼 polymerizing에서 G - 굴지 흐르는하여 챔버 내부의 중합 굴지.
  5. 초과 버퍼 흐르는으로 희미 unpolymerized의 굴지.

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Comments

1 Comment

  1. What is the second pair of shorter parafilm strips for? Why do you polymerize in the chamber and how? G-actin in polymerization buffer. Is't that F-actin? What is the actin binding agent and blocking protein and their buffers and their concentrations?

    Reply
    Posted by: Anonymous
    July 28, 2009 - 2:41 PM

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