- I think the original poster was only calling attention to the fact
that some proteins want to be treated respectfully in order to
crystallise (and the fact that Rigaku Japan realises this). I find
that indeed the case.
Other proteins, however, prefer the attitude "I don't know why I am
setting up these drops, this protein is too crappy to crystallise",
i.e. a challenge.
Lysozyme, on the other hand, even crystallises under conditions of
complete indifference. At least I find that every student in a
practical course can get nice crystals of lysozyme, and a majority of
these drops have been set up under conditions of complete
indifference...maybe lysozyme is not a protein after all, but a salt:
lysozyme-chloride / LyCl7 ?
Mark
PS the detailed protocols and experiences are useful though.
Quoting Jeffrey Wilson <wil...@ucmail.uc.edu>:
I was recently testing out the 30% PEGMME 5k, 0.1M NaOAc pH 4.5, 1M
NaCl method mentioned in a Hampton Research catalog and attributed to
Enrico Stura. I see that he has also just commented on this thread. I
found that at 80mg/ml, batch with 1.5ul:1.5ul protein to precipitant
ratio, lysozyme crystallized in about 1 hour. Jumping that up to
150mg/ml allowed for crystallization in minutes. Hanging drop behaved
similarly. I was using lysozyme from Sigma.
Jeff
Jeffrey Wilson, Ph.D.
University of Cincinnati College of Medicine
Molecular Genetics Department
231 Albert Sabin Way
MSB 3109A
Cincinnati, OH 45267-0524
(513) 558-1360
On Dec 21, 2009, at 11:12 AM, MARTYN SYMMONS wrote:
Dear All
checking out the Lysozyme crystallization methods on the web I
liked the Rigaku Instructions that I found:
(http://www.rigaku.com/protein/crystallization.html)
"...create a drop of 3ul lysozyme solution, and 3 ul of well
solution, respectfully, for a total drop size of 6ul..."
So perhaps sometimes I am just not respectful enough to deserve crystals ?
good wishes to all
regards,
Martyn
-----------------------
Martyn Symmons
MRC-MBU Cambridge UK
'Chan fhiosrach mur feòraich.'
Gaelic proverb -
Nothing asked, nothing learned.
