streaky page gels

timothy patrick purk purk at
Mon Jul 8 20:28:12 EST 2002

Hi again,

I am still having trouble so let me give you alittle more information
about my problem.  I have done a couple rounds of IE chromatography on
Xenopus egg extracts and am running the fractions that were positive by
western over a Superose6 size exclusion column.  My protein is quite large
so I am running a straight 6%gel with no stacker as my protein seems to
get hung up at the interface when I include a stacker.  After the SE
chromatography, I am TCA precipitaing the fractions and resuspending in a
standard 1X loading buffer.  The prestained markers look fine and by
looking at the dye front, I don't see anything weird about how the gel
is running.  I am blotting with the same Ab that I used to verify the
presence of my protein in previous steps with  the same conditions.
However, when I run these samples, all I see after developing my western
are linear streaks running straight up and down where my lanes are and no
detectable bands.  Thanks to everyone who has written so far, I am
completely flumoxed by this, and am ready to throw the column out.

T. Patrick Purk

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