Multiphor 2-D electrophoresis

Litta Olsen litta.olsen at sars.uib.no
Fri Mar 27 03:50:47 EST 1998


Hi.
For some time now I have been trying to make the 2-D system from
Pharmacia work and have some trouble with this.
I use premade Dry Strips and precast gels (8-18%)
The first problem was that the front in the 2. dimension would move much
faster on one side than on the other. This problem has been reduced, but
still I see a smiling effect on one side, and since I do two parallel
strips on one gel in order to compare them, this makes it rather
difficult.
I started out using the ph3-10 strips, but realised that they did not
separate my proteines well and decided to try the pH4-7. I did the 2.
dimension yesterday and it gave a strange pattern. It seems to me that
all the proteines had "jumped" over about 1/3 of the strip.
Approximately one third of the strip in the pH7 end was empty and it was
not like it gradually dissapeared.There was a clear vertical split
between the areas with an without proteines. I am sure this is due to
the 1.dimension run, since both strips gave exactly the same pattern. 

Has anyone an answer to this, then please contact me.

Thanks
Litta



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