Dear Lu,
To be quite honest, before running refinement, I still use a simple program to
add and delete waters based on electron density and distance criteria, which I
wrote a long time ago in the dark ages of Vax computers and Evans Sutherland
graphics when automatic water update did not
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
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
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,
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
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 rmgarav...@gmail.com
wrote:
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
Apparently these are not for sale anymore by EMD Millipore
yet it is what is recommended for plasmid preps with pETcoco-2.
Does anyone know of a source of these cells
or a substitute cell line we could use?
Thanks, Gloria
Dear all,
I'm sorry, this is a bit off-topic. I'm looking for a tool to compare pairs of
binding sites.
It doesn't need to be high-throughput, since I'll only be comparing ~100 pairs,
but I'd like it to be robust and, more importantly, alignment-independent.
Indeed, the binding sites I want to
Dear Sarah,
we don't have a server for you to quickly use but our lab has methods for
alignment-free binding sites comparison by comparing pocket surface shape,
named Pocket-Surfer and Patch-Surfer.
These are literature:
http://www.ncbi.nlm.nih.gov/pubmed/20455259
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