By size exclusion, it eluted where a 30kDa protein would be expected to elute.
As for functional assay, I am currently trying to express its binding partner,
though it shows a lot of degradation. But when I mix both proteins and run by
size exclusion it causes the peaks of its binding partner to
Hi Dan,
I agree with Bert's general approach. How does your purified
protein look under size exclusion chromatography or DLS? Do you have a
functional assay?
ho
It behaves very well. No precipitation/cloudiness during purification.
Secondary structure estimation by CD is ~50% alpha helical, 25% beta-sheet and
25% random coiled. I screened at 2, 4, 6 and 8mg/ml (8mg/ml around about
60:40% precipitation). If any other information is needed, just ask.
Th
Hi Daniel,
Whether or not I would introduce detergents would depend on the behavior of the
protein during purification and crystallization. Does it behave well during
purification in the absence of detergents? In your crystallization screens, do
most of the drops have heavy precipitation? If th
I have a 30kDa protein which I have been trying to crystallize. I have tried in
the conventional way using Hampton screens but no luck so far. CD of the
protein shows it is folded. However reading the literature I found out it is
strongly associated with the inner membrane of H. pylori. This was
Hi,
You must have a special reason for doing what you plan at this stage, although
it's definitely not clear to me why. Refmac5 deals properly with twinning and
the output is ready for deposition as pdb, including the twinning information.
In addition, the ccp4 package has also options to deal
Hi Christine
Do you know the conditions for crystallization of the Fab only ? You might
start using the Pro-complex screens to start with and then use DMSO
solubilized peptide in and around that solution.
Thanks,
Ivan
On Mon, Jul 19, 2010 at 10:37 AM, Harman, Christine <
christine.har...@fda.hhs.
