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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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- Construct a flow chamber as shown in the video. Make sure that the parafilm seal is tight and use washed coverslips.
- Incubate actin-binding agent in the chamber for 5-10'.
- Block non-specific binding sites on the glass coverslip with a blocking protein.
- Polymerize actin inside the chamber by flowing in G-actin in polymerizing buffer.
- Washout unpolymerized actin by flowing in excess buffer.