Surveillance actine démontage Time-lapse microscopie

Published 11/08/2006
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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. Construire une chambre d'écoulement comme le montre la vidéo. Assurez-vous que le sceau du parafilm est serré et l'utilisation des lamelles lavées.
  2. Incuber actine agent liant dans la chambre pour '5-10.
  3. Bloc non spécifiques des sites de liaison sur la lamelle de verre avec une protéine bloquant.
  4. Actine polymérise intérieur de la chambre par l'écoulement de G-actine à polymériser tampon.
  5. Délavage polymérisés actine en s'écoulant dans l'excès de tampon.

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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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