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Isse with "Smiles" on gel

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#1 poust



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Posted 22 January 2010 - 01:26 PM

I'm attempting to extract the outer membrane porin OmpF from E. Coli and I seem to be having trouble. The overall extraction protocol is here Protocol. The basic version is that I am attempting to extract OmpF from an outer membrane pellet using the detergent Octyl-POE. I'm wondering if the smiles on these gels is due to: too much detergent being loaded on the gel in my sample, some other SDS-PAGE problem or another problem with in my purification. My ladder is running fine so I don't think the problem is with the gel. Please take a look at these gels and see what problem this looks like to you. The "smiley" bands in these purification gels are my final extraction supernatant samples, which are in 3% Octyl-POE.

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Thanks! <_<

#2 mdfenko


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Posted 22 January 2010 - 09:33 PM

either the detergent or the lipids they solubilized are causing the distortion.

try dialyzing the detergent away from the sample before adding the loading buffer.
talent does what it can
genius does what it must
i do what i get paid to do

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