membrane protein extraction - membrane protein extraction (Jul/02/2008 )

Hi all,
I'm trying to extract membrane proteins( from rat cardiomyocytes) for western blot analysis. So far I have tried Triton X 100, NP-40, SDS and still I dont have any satisfactory result. Could anyone please suggest something to try out.
Thanks,
Tulip

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Tulip, while some suggestions come along, check here if you find some good method. Let us know once it works!
http://search.vadlo.com/b/q?sn=158621799&a...ction&rel=0
...

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Thanks a million, I will surely get back once I try some of these methods.
Tulip

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Try adding deoxycholate to your extraction buffer, it is a very strong detergent (check out RIPA buffer).

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Try AquaRNA. It's a mix of potent chaotropic agents and detergents, in addition to specific RNase inhibitors.

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Thanks for your suggestion, I tried 0.5% deoxycholate along with 1% Triton X 100 too, but I'm still not getting complete solubilization of the membrane fraction.

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Has anybody tried Triton X 114 phase partitioning method?

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May be you should try nonionic surfactants
such as C11EO2(Berol 532), C9-11EO5.5....
they gave been used to purify membrane
proteins for a long time.
gud luck

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