Monitoreo de actina desmontaje con la Microscopía de Time-lapse

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. Construir una cámara de flujo como se muestra en el video. Asegúrese de que el sello de parafina es escaso y el uso de cubreobjetos se lavan.
  2. Incubar actina agente fijador en la cámara para '10.5.
  3. Bloquear no específicas sitios de unión sobre el cubreobjetos con una proteína de bloqueo.
  4. Actina polimeriza dentro de la cámara de flujo en G-actina en la polimerización de amortiguamiento.
  5. Lavado de actina polimerizada por el flujo en exceso de tampón.

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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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