Hi Leonard,
I would treat this like a MS/MS ion search experiment looking for the peptide
via MASCOT or similar. Talk to some local Mass Spec people, I'm certain they
could identify it and then you'd be closer to an explanation.
John.
Dr. John Hinks
Syngenta
Jealott's Hil
Hi Yang.
Adding to what Phoebe has said below, remember that when running polyacrylamide
shift gels using TBE and in the absence of SDS, your protein (depending on its
charge) may not migrate into the gel, or may in fact migrate out of the well
altogether (in the absence of DNA binding).
In or
Hi Ganesh,
two things I'd advise that might make a difference for you:
1. Improved media for better expression
See "Protein production by auto-induction in high-density shaking
cultures" by W.F. Studier Protein Expression and Purification 41 (2005)
207-234
(there's also a re
Hi Matt,
if you have a crystallisation robot (and if you haven't you can still do this
manually) you can use a simple microbatch under oil experiment to determine the
likely optimum pH and salt concentration.
Just set up a grid across pH (I usually use a phosphate buffer for this as it
is easi
