2d gel cleanup

Nick Theodorakis nicholas_theodorakis at urmc.rochester.edu
Fri May 19 16:49:35 EST 2000


In article <39258CD6.9815A5D2 at uiuc.edu>, Andrea Beckel-Mitchener
<
amitch at uiuc.edu> wrote:
>I am new at 2D gel electrophoresis and would appreciate some
technical
>advice.
>
>I have synthesized proteins (labeled them with 35S-Met) and run
a 2D
>gel.  There is quite a bit of background from the radioisotope,
yet I am
>able to see very good "spots" as well.  I would like to clean
up the
>appearence of the gel by possibly getting rid of unincorporated
label if
>possible  (much of the signal runs with the dye front and
doesn't pose a
>problem, it's the rest of it that is annoying).  Can I
precipitate
>proteins and still run a good 2D?  I was concerned that
precipitation
>with TCA might interfere with running the IEF dimension.  Any
>suggestions/advice would be great.  Thanks.

Are you fixing the gels beore drying them? Usually, fixation and
washing should wash out the unincorporated label.

Nick


Nick Theodorakis

nicholas_theodorakis at urmc.rochester.edu
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