Very very mysterious SDS-PAGE - (Jan/29/2007 )

Hi all,

I have a very strange problem about the SDS-PAGE. Recently, when I ran the gel on one side of the eletrophoresis cell, the proteins were all degradated... But this doesn't happen on the other side of the cell. This is very weired because I made, poured and ran the two gels all together, the reagents and samples were all the same, but the degradation of proteins only happened on one side of the cell and the gel on the other side seems nice and normal... This had happened not only to me but also to other members in the lab.
Has this occured to anyone? Does anyone know what on the earth made this happen??

Thanks!
Elaine

-Elaine Chou-

This hasnt happened to me.

I would guess something is not right with the chamber. Could be some electrical connectivity problem. Have you or others tried a different chamber and a different lid?

-scolix-

QUOTE (Elaine Chou @ Jan 29 2007, 07:08 PM)

degradation in the sense of proteolysis? there may some contamination of proteases as part of the sample preparation (intrinsic proteases) or f.i. from ungloved fingers (extrinsic proteases); decontaminate your gel electrophoresis equipment carefully

-The Bearer-

Actually, the samples for the two gels were the same, so I don't think it's the problems of my sample preparation...
I tried to use a different lid, and this still happened. I think it must be the problem of the chamber...

-Elaine Chou-

"Could be some electrical connectivity problem"
I would think so too. not sure how your cell was set up
but recently I found one part of my gel apparatus heat up while the other did not
(I didn't run any sample on that side though)
and then I found that the "electric wires" had come off and has not been going through its right track.
Anyway, it would be good to check that part a little
good luck!

-Lynnpanda-