Smeared Western Samples

Sulakshana Mukherjee mukherji at tifr.res.in
Mon Nov 12 08:09:49 EST 2001


Dear Sophie,
I have experienced a similar problem. I fixed the problem by simply
changing the gel-running buffer ( by making fresh buffer).

with best regards,
Sulakshana

On Mon, 12 Nov 2001, Araluen Freeman wrote:

> 
> "Sophie" <sweiss at icr.ac.uk> wrote in message
> news:20011109105429.9175.qmail at ww02.jatek.com...
> 
> > I am trying to run western blots to determine the level of protein
> > expression in salmonella. I lyse the bacteria with 1% triton, 10% glycerol
> > and 15 aprotonin. My protein is 46000Da and i run it on a 12% SDS PAGE
> gel.
> > The markers are always nicely separated and transfer to the membrane.
> >
> > My problem is that all my samples end up as horrible smears down the gel
> > instead of compact bands. I thought it was a problem with the bacteria but
> > the same thing happens when i use a highly purified version of my protein.
> > Has anyone got any suggestions for why this is happening and how i can fix
> it.> 
> 

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