Monitoramento Actina Desmontagem com a Time-lapse Microscopia

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. Construir uma câmara de fluxo, como mostrado no vídeo. Certifique-se que o selo parafilme é apertado e use lamínulas lavadas.
  2. Incubar agente de ligação de actina na câmara para '5-10.
  3. Bloco não-específicos sítios de ligação na lamela de vidro com uma proteína de bloqueio.
  4. Polimerizar actina no interior da câmara de fluxo em G-actina na polimerização buffer.
  5. Washout actina unpolymerized fluindo em tampão em excesso.

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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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