Problems w/ native PAGE

[POPOVA]

Hello,

>I have been having problems with native PAGE of proteins.  When I extract
>total proteins from tobacco flower organs or leaves (I've listed my
>protocol below) and run them on 4 to 20% denaturing SDS gels, I get fine
>resolution of bands with no detectable smearing.  If I run the same
>amount (30 ug) on a 15% native gel, the bands are poorly resolved and
>smeared along the edges of each lane. 

DEAR PAUL,
I WOULD TRIED THE FOLLOWING:
1. DISSOLVE  PROTEINS BEFORE PAGE IN 8M UREA
2. MORE EFIICIENT COOLING (COLD ROOM?)
3. DECREASE (WHERE APPROPRIATE) THE LOADING PER LANE

SOMETIMES I INCLUDED UREA INTO THE GEL AS WELL

HOPE THIS HELPS
ANDREI POPOV