Monitoring Actin Demontage met Time-lapse Microscopie

Published 11/08/2006
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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. Construeer een stroom geplaatst zoals aangegeven in de video. Zorg ervoor dat de Parafilm afdichting strak is en het gebruik van gewassen dekglaasjes.
  2. Incubeer actine-bindende middel in de kamer voor 5-10 '.
  3. Blokkeer niet-specifieke bindingsplaatsen op het glas dekglaasje met een blokkerend eiwit.
  4. Polymeriseren actine in de kamer door stroomt in G-actine in polymeriseren buffer.
  5. Wash-out niet-gepolymeriseerde actine door stromend dan buffer.

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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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