Övervakning Actin demontering med Time-lapse Mikroskopi

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. Konstruera ett flöde kammare som visas i videon. Se till att parafilm tätningen är tät och använda tvättad täckglas.
  2. Inkubera aktin-bindemedel i kammaren för 5-10 ".
  3. Blockera icke-specifik bindningsställen på glaset täckglas med en blockerande protein.
  4. Polymerisera aktin inne i kammaren som flyter i G-aktin i polymerisation buffert.
  5. Washout Opolymeriserat aktin som flödar över buffert.

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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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