need a good protocol for actin staining - (Dec/07/2007 )

We're using phalloidin to detect actin in tissue samples, at least, that's the idea. I'm having problems with the protocol we have, and did not find many online. Does anybody have a good, solid, working one?

Thank you

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search the protocol section of this website and you will find some protocols that you may be able to adapt.

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Thanks for the answer. I already have -and found papers with phalloidin staining, but I was hoping somebody has a magic bullet, that solves all my problems

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I have done that on tissue sections. I got good results without much troubles. Liver samples by perfusion fixation with 4% PFA, croysection to 8 um.
Its a simple incubation for up to 30 min, washing several times, that is it. I got the FITC-conjugated form from sigma. You can use better ones like Alexi-labeled for longer photostability. I used the dilution suggested. Not thing spacial. Only that its been quite years back, and we never publish that work. I can't give you exactly what did I do. So tell us what problems do you have and maybe we can help you.

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I confirm that Alexa-fluor-labeled phalloidin (Invitrogen) is better than FITC-labelled; we use the same protocol as for antibody staining

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Thank you all for the answers. We have paraffin embedded tissue. Do you think that paraffin is incompatible with actin staining? I can not find any protocol for paraffin embedded tissue, and your protocols do not perform very well with paraffin...

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Maybe some of these discussions will interest you?

http://search.histosearch.com/search.acgi$search

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I'm sorry, but the link does not lead anywhere. Would you please post it again?
Thank you

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Sorry, I used the following link and typed "actin" http://www.histosearch.com/histonet/

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Thank you, I'm combing through it (So far I found a post with the same question that no one answered...)

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