Hello,
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.
Thank-you
0
1 reply to this topic
#2
mdfenko
-
- Active Members
-
- 2,286 posts
an elder
83
Excellent
Excellent
Posted 22 February 2012 - 01:04 PM
the smearing could be due to post-translational modifications on the protein (eg glycosylation, lipidation).
it could also be caused by components in the sample's buffer.
it could also be caused by components in the sample's buffer.
talent does what it can
genius does what it must
i do what i get paid to do
genius does what it must
i do what i get paid to do
