Dear Rafael,
do you know if your protein interacts with another protein? This could be
critical. I have been studying proteins that require a protein partner to be
expressed as soluble protein.
Co-expressing both proteins in E. coli shifts the expression from insoluble to
soluble.
Hope this h
Hello,
I found this one, 56 minutes but the quality leaves much to be desired:
https://web.archive.org/web/20180304013635/http://www.thirteen.org:80/naturally-obsessed/wp-content/blogs.dir/30/files/2013/01/Naturally_Obsessed_The_Making_of_a_Scientist.mp4
Best regards,
Oleg Sobolev.
On Tue, Mar 1
Hello Rafael,
I would give one more by reducing the temperature for the expression. I am
assuming that you did the expression at 37 oC. I would start expression at 30
oC and reduce it further after induction to 25 oC or so and let it grow longer.
But refolding the protein is also possible. T
The problem is not to ensure confidentiality of what one writes. It is to
ensure that one can write publicly (within some limits, see below) without
fear. To go back to the original conversation, I think we have here an example
of a “pendulum effect”. The onslaught on what cannot be said
Dear colleagues,
I would be most grateful if you could bring the opening below to the attention
of potential candidates.
We have an opening for a 50% Technical Assistant to help with all tasks of
laboratory maintenance, management and stocking. Familiarity with the
techniques applied in a pr
Hi Dave, I should have mentioned that, but yes, I tried 16 degrees overnight and even up to 40 hours. The _expression_ level is great, majority is in the pellet though and the elution fraction is far from desirable for my kinetic assays. Best wishes _
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I guess I should remind subscribers that the CCP4BB has a public archive,
at https://www.jiscmail.ac.uk/cgi-bin/webadmin?A0=ccp4bb.
On a personal note, I always appreciate the courage of those who call out
the bullshit in the world, and I've enjoyed the eloquent messages in this
thread (though the
Then: on the poop list of the postmodernists:
Transgressing conventional boundaries: Postmodern Structural Biology. * B.
Rupp. IUCR Keynote. Aug. 21-29, 2011*
Now: on the poop list of the new order:
Correcting cis-trans-transgressions in macromolecular structure models. F.
Waibl, K. R. Liedl, B. Ru
These discussions remind me of a talk I heard from Shekar Mande, I think, about
the role of crystallographers in the wars around the world, particularly how
this community has stood up for fairness.
I have never felt prouder to be part of this community. I believe that as
scientists, we also ha
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