Smeared Western Samples

Sophie sweiss at icr.ac.uk
Fri Nov 9 05:54:29 EST 2001


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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