1 reply to this topic

[poust]

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.





Thanks!

[mdfenko]

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.