Why has does my developed blot look smeared? (Western Blotting) - Western Blotting Analysis (Mar/15/2007 )

Hi there...

I made up and ran my gel (7%) as normal...nothing seemed out of the ordinary when i transferred (molecular markers came through nice and bright like i'm used to seeing!)... and then when i developed the blot, i see a whole lot of smeared lanes.... predominantly around the middle. i re-'ECL'ed it but had the same result.. when i asked my supervisor, he seems to think (as did I) that it was something to do with the gel - that it hadn't set properly? but he's also not sure (says he can't remember!) is that what happens if the gel smears? but then, my markers have come out fine... i don't get it... never had this problem before....hmmm...


Archana

you may have overloaded your samples.

Hi Archana
It is magbe the overloaded samples or your samples contain more salt.

Hi there...

I made up and ran my gel (7%) as normal...nothing seemed out of the ordinary when i transferred (molecular markers came through nice and bright like i'm used to seeing!)... and then when i developed the blot, i see a whole lot of smeared lanes.... predominantly around the middle. i re-'ECL'ed it but had the same result.. when i asked my supervisor, he seems to think (as did I) that it was something to do with the gel - that it hadn't set properly? but he's also not sure (says he can't remember!) is that what happens if the gel smears? but then, my markers have come out fine... i don't get it... never had this problem before....hmmm...


Archana