How to research on acin? - (Oct/26/2006 )

Recently, i got a protein which interact with actin. The further research on it needs function research of the interaction, as you know, actin is familyhold gene, so knockout of the actin gene is impossible. anybody can guide my research to deeper. Reply would get my appreciates.

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your further approaches depend on the kind of protein you have; or is it totally unknown? to knock out actin won´t do but there are pharmacological agents such as cytochalasin D which disrupt F-actin filaments, or others such as jaspakinolide promote actin polymerization; you may follow if your protein binds also to G-actin or will only bind to polymerizing or polymerized F-actin

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Thank you, i have read many articles about actin, in which cytochalasin D is a common tool for research on actin, and you gave the same pharmacological agents, therefore i plan to use it.
another question is that recent research on actin make use of confocal microscopy, actin filament turnover assay, or cell motility assay, i want to get deep sight in the research ways, any articles about it can help me.

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for CLSM you can perform life cell imaging, therefore a fluorescent protein-tagged actin is useful, you may check Clontech now line of Takara I think; if you try FRET think of CFP-YFP or GFP-red FP which should influence your decision which fluorecent dye your actin should have;

for ICC/IHC, you may use phalloidin-Alexa XXX, we use 633 as we use the other channels for 488 and 546 labelled secondary Ab, or also fluorescence tagged actin to express

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