isolation of nuclear protein - isolation of a nuclear protein, namely Akt protein (Mar/10/2005 )

Hi all,

I would like to isolate Akt protein, a kinase, from nuclei of cardiomyocytes. Do any of you have experience in this topic ? Please give e.g. a standard protocol for isolation. I should use it at first in western blott.

Thank you for information.

Andree

hi
i tink you first have to do total proteins nuclear extract.
You can find a protocol here :

http://www.biochem.northwestern.edu/ibis/m...%20Extracts.pdf

here is a receipe for cytoplasmic and nuclear extract :
http://www.protocol-online.org/cgi-bin/prot/jump.cgi?ID=1534

finally here the global nuclear extract page from protocol online :
http://www.protocol-online.org/cgi-bin/pro...&submit2=Search