Cell Envelope Proteins Smeared
Posted 22 February 2012 - 09:46 AM
I am attempted to do a western on cell envelope protein I have isolated from P. aeruginosa, however the lower proteins always come out smeared. I have ran gels that are 10%, 12% and 15%. I store my protein in -20 with PMSF, I run at 80V for 30 min then 120 for 2.5 hours. My ladder is always clear. I have remade all of my buffers and have changed my sonication time and amplitude. I am at a loss.
Any input would be appreciated.
Posted 22 February 2012 - 01:04 PM
it could also be caused by components in the sample's buffer.
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