In my lab we are still making the old fashioned tube gel IEF.
We are having problems with the tube gel polymerization, after 2 hrs of polymerization, the gel just drain out of the tube.
Everything is new but the ampholytes. I was wondering if is a matter of urea solubility.
Can anyone help me with this?
Thanks!
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#2
mdfenko
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Posted 02 March 2010 - 01:04 PM
how old are the ampholytes?
when did you last use them?
are you sure bisacrylamide (or other crosslinker) was added to the acrylamide solution?
you can confirm that it is not the fault of the ampholytes by adding a little temed to the gelling solution and seeing if it polymerizes. if it does then it is probably bad ampholytes. if not then it is either the acrylamide or the persulfate solution that is the culprit (try a different batch of persulfate)
when did you last use them?
are you sure bisacrylamide (or other crosslinker) was added to the acrylamide solution?
you can confirm that it is not the fault of the ampholytes by adding a little temed to the gelling solution and seeing if it polymerizes. if it does then it is probably bad ampholytes. if not then it is either the acrylamide or the persulfate solution that is the culprit (try a different batch of persulfate)
Edited by mdfenko, 02 March 2010 - 01:08 PM.
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#3
Ivanov_br
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Posted 08 March 2010 - 12:08 PM
mdfenko, on Mar 2 2010, 06:04 PM, said:
how old are the ampholytes?
when did you last use them?
are you sure bisacrylamide (or other crosslinker) was added to the acrylamide solution?
you can confirm that it is not the fault of the ampholytes by adding a little temed to the gelling solution and seeing if it polymerizes. if it does then it is probably bad ampholytes. if not then it is either the acrylamide or the persulfate solution that is the culprit (try a different batch of persulfate)
when did you last use them?
are you sure bisacrylamide (or other crosslinker) was added to the acrylamide solution?
you can confirm that it is not the fault of the ampholytes by adding a little temed to the gelling solution and seeing if it polymerizes. if it does then it is probably bad ampholytes. if not then it is either the acrylamide or the persulfate solution that is the culprit (try a different batch of persulfate)
Thanks for your reply! I'll try a new ampholyte this week! I'll post the news!
