protein extraction with phenol/SDS - (Jun/06/2008 )

I want to extract proteins from leaves of ressurrection plant Ramonda serbica, which has a lot of phenol and other interfering compounds. I used method with dense SDS buffer (succrose, SDS, Tris-HCl, and beta merc.ethanol) and phenol. When I precipitate my proteins with ammonium acetate I have pellet but I have problem to resuspend it in Laemmli buffer. Can I use sonification?Does anybody know some buffer which I may use? maybe buffer with Tris-Cl and NaCl, but in this case I have to add PVP?Does anybody know?
Zix

-zixoni-

QUOTE (zixoni @ Jun 5 2008, 11:38 PM)

Not a specific answer, never worked on plant projects, but you may find some good protocols here.
http://search.vadlo.com/b/q?sn=158621799&a...ction&rel=0
..

-cellcounter-

i work with plants but not with proteins.... however i think sonication might affect your proteins but don't trust me too much on that. i've been with maize endosperm to extract RNA, lots of interfering compounds as well. the advantage i had was using columns. maybe increasing your buffer volume could help resuspending the pellet?

-toejam-

Thank's! I have tried with Tris-Cl,NaCl,PVP40 and Triton X100 and there was no problem.

-zixoni-

hey.. there s trizol reagent and tri reagent.. u can isolate protein using those...

http://www.cosmobio.co.jp/product/products...18.20060623.pdf

this should help..

-krithika-

hey.. there s trizol reagent and tri reagent.. u can isolate protein using those...

http://www.cosmobio.co.jp/product/products...18.20060623.pdf

this should help..

-krithika-