Thank you all for your replies.
I tried seeding also. But it did not help me.
I ran SDS-PAGE. The fresh protein and crystal are showing bands at same
molecular weight. So there might not be protein degradation.
On 3 November 2014 22:24, Appu kumar wrote:
> Hello,
> If the homologous protein st
Hello,
If the homologous protein structure is available, or you have symmetry
information of your crystal, then try engineering surface by putting in
residues which has less entropy like replacing Lys to Arg, Met to Gln. We
are able to fasten the crystal growth from 20 days to 3-4 days by creating
Michael (and some others)
You haven't mentioned - and I guess don't use regularly - the "random" MMS
approach, where crushed seed-crystals are added to random screens. This
really is a terrific method, and it will on average roughly double
productivity. It's the first thing I would think of in a
I assume you were using vapour diffusion. In that case, you could also try
microbatch to cover the possibility of the plate slowly drying out and
concentrations increasing beyond those of the original screen.
Dmitry
On Fri, Oct 31, 2014 at 12:07 PM, R. M. Garavito
wrote:
> Although three months
Although three months is a long time, it is no completely unheard of, and does
not require the invocation of proteolysis. The longest time I have heard of is
~1 yr, so count yourself lucky. However to get good advice, as well as to use
it, you need to ask yourself several questions:
1. What
Dear,
the conditions you list should equilibrate within a couple of days. A
growth time of three months could be a sign of some proteolytic reaction
happening. You might want to sequence the crystal and subclone the
fragment that actually crystallises.
Best,
Tim
On 10/31/2014 11:05 AM, Vijaykuma
I recommend to try seeding!
On 31.10.2014 14:05, Vijaykumar Pillalamarri wrote:
Dear all,
I am trying to crystallize a 30 kD protein. Protein crystals are
formed after 3 months. The crystals are formed in the following condition:
0.01M Zinc sulphate
0.1M MES monohydrate pH 6.5
25% v/v PEG mono
Dear all,
I am trying to crystallize a 30 kD protein. Protein crystals are formed
after 3 months. The crystals are formed in the following condition:
0.01M Zinc sulphate
0.1M MES monohydrate pH 6.5
25% v/v PEG monomethyl ether 550
Please suggest me how to grow these crystals faster.
Thanking you
