SDS gel strangely running: curved ladder - Ladder starts running vertical, and then levels horizontal in the running gel (Feb/01/2006 )
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Hi everybody,
My westernblots protocol has been the same for 2 years now, also other people on the lab are using this protocol, but since a few weeks I have this problem.
My ladder is curved, it even starts to level out horizontally.
The strange thing is that in only happened on one gel in the mini-blot system and not on the other! (with samples of my colleague)
Can it depend on our different isolation buffers?
I used this buffer quit time without problems.
I made the gels exactly on the same moment, and waited for 40 min before adding the stacking gel.
hi
may you join a picture of your gel?
For my exp, i have sometimes problems with te ladder if it's on the first lane of the gel (or the 10th of a 10lanes minigel). Starting horizontal and slowely the inner side of band is downer than the outter part of the band... 
but i don't know why.
idea : when i clean and dry with ethanol the stuff for the gel (glass spacers and the comb) i get nice migration.
Hello,
It might be due to a high concentration of salt (in the samples).
how's your glycerol concentration?
how's the pH of your buffer?
I think that the platinum (the metal that moves the current) is probebly broken. this is why it only happens on one of the mini gels.
either fix it your self or send it to someone to fix it for you
good luck 
either fix it your self or send it to someone to fix it for you
good luck
Reply from Eveline: No, the metal wire is not broken, that is the first thing we checked.
Reply from Eveline: The ladder was in the 4th lane, I usually do not use the first lane because of what you said.
I do not have a picture, cause I threw the gel away, but the ladder on my side looked like this:
0____
0____
00000___
000000000__
00000000000000 _
Strange, don't you think?
I am going to make a new isolation buffer.
Maybe someone contaminated the old iso-buffer.
The pH of the buffer is supposed to be 7,5