رصد أكتين التفكيك مع مرور الزمن الميكروسكوب

Biology

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Summary

Cite this Article

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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. بناء غرفة تدفق كما هو مبين في الفيديو. تأكد من أن الختم parafilm ضيق واستخدام coverslips غسلها.
  2. احتضان أكتين ملزم عامل في غرفة ل '5-10.
  3. كتلة غير محددة على مواقع الربط ساترة الزجاج مع بروتين حظر.
  4. تبلمر أكتين داخل الغرفة التي تتدفق في G - أكتين في البلمرة العازلة.
  5. تبييض unpolymerized أكتين العازلة التي تتدفق في الزائدة.

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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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