Weird SDS-PAGE... HELP!!
jv4s at avery.med.Virginia.EDU
Tue May 9 14:12:29 EST 1995
In article <95128.132452JPS144 at psuvm.psu.edu>,
Joe Stains <JPS144 at psuvm.psu.edu> wrote:
>Someone in my lab is running SDS-PAGE on a cell lysate. However, when
>they run the gel, the sample lanes spread out (longitudinally), pushing
>the adjacent lanes out as well. No one seems to have seen anything like
>Among the steps we've done are: 1) run samples containing positive
>control samples with added lysate buffer => no problems with the run 2)
>4) running gel at lower amp (20mA constant)
Whenever I run cell lysates on gel I make sure there are no empty lanes
on the gel. I've noticed that empty lanes encourage lane spreading at the
bottom. I fill the empty lanes with a similar dilution of sample buffer.
This seems to help.
The other problem may be heating of the plates. Running mine at 20mA
(we run laemmlis) dont seem to warm them much so I'm not
sure if that is your problem
If you overload the lanes with protein you shouldn't have a lane spreading
problem, your bands will look like crap though!
Jayanand Vasudevan (804) 982-1891
Dept. Pathology/Biochemistry, Box 214,
UVa Health Sciences Center Charlottesville VA 22908
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