I can't remember if someone has already suggested this. You can
dissolve some of your crystals and ask your favorite mass spec. lab to
check if your protein has been oxidized, proteolyzed, etc.
ho
Confometrx
Dear James --
One possible issue you might have as well is oxidation. Your sample
might need to be in a different step... and would therefore require
some time before getting to this step. I personally noticed (in few
cases) that when my crystals took time to grow, it was either because
t
Sometimes it is better that it takes time for crystals to appear.
Remember that crystallisation is a purification procedure. A way to
decrease the speed of crystallisation is to use Dunlop's and Haze's drop
dilution method (K. V. Dunlop & B. Hazes (2003). When less is more: a
more efficient vap
Hi James,
have you tried limited proteolysis on your protein and see if you can
identify a stable fragment. Then re-clone and re-crystallize your
protein. Or a very stupid suggestion, how does your size exclusion
peak look like ? What you're not running your protein over a SEC to
polish i
I've always found it is a much more effective and delightful use of my
time to mope sullenly in my office, communicate with my family members and
colleagues while restricting my working vocabulary to monosyllabic grunts,
or pensively pace the halls, inflicting my abject misery upon hapless
passers-
depending on the importance of the project, I would be tempted to just
wait for the crystals to grow, and in the meantime:
- work on another project
- start a new project
- write a grant application, a paper, a review...
or a combination of the above.
Mark
Quoting William Scott :
Dear James:
It will help more if we send it to James ;)
On Oct 11, 2009, at 9:15 AM, gauri misra wrote:
Dear James,
As there are indications of protein degradation that have been
suggested in
previous postings, i think adding some protease inhibitors right at
the
stage of purification may provide so
Dear James:
It is extremely hard to make generalizations, but if it takes months for
crystals to appear, the following questions are worth asking:
1. Is the actual concentration of macromolecule, precipitating agent, etc
required for crystallization higher than what you are using? A closed
vapor
Dear James
What size are your drops? If it takes that long for crystals to grow, it is
very likely that something happens to the protein that stopped nucleation
previously... it generally means some form of protein degradation. I would
start by investigating exactly what you have on the crystals. I
Dear crystallographers,
Sorry for the non-ccp4 query. I am new to this field and need some
suggestions. My question is, why some protein takes longer time to
crystallize, say 6-8 months, and it is the only condition to get the
crystals.? What are the ways to get the crystals faster.
The crystal a
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