Is there a method/protocol to stain nuclear actin by, e.g. phallodin? I cannot afford fused ABDs-fluorescent proteins now, is there an alternative?
Phalloidin works, but be careful with your fixative. RT Methanol destroys actin, and formaldehyde or paraformaldehyde kill it with a half-life of about 15'.
Thorough permeabilisation is also important.
I expect you could get someone to give you an ABD construct for free...
It is quite hard to visualize actin in the nucleus because the filaments are much thiner there. Actually for years, people thought there was no actin in the nucleus because it could not be visualized. Permeability after fixation is key. I would recommend taking Dr. Insall's suggestion and requesting the construct you need from someone.
Hello
visit this link
http://www.cytoskeleton.com/actin-staining-techniques
Hope help you .Good Luck
Hi Dmitry,
ABPs are ok but be careful as 1) different ABPs mark different nuclear structures, and 2) they may also stabilise F-actin thereby creating (ABP-specific) artefacts.
I would first try with phalloidin and only later employ ABPs.
Good luck with your experiments!
Metello
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