Ji,
probably the oil doesn't work and the glycerol gets you into problems
with phase separation in 2.5 M AmS
I recommend sucrose or other sugars with AmS - 20-22% (w/vol) should
do and you get no phase separation
At the same time you may need to increase the AmS considerably - often
the pro
Thank You very much for all your suggestions and reply. i would consider all
of this to sort the issue. once again thanks a lot.
On 6/23/08, Exec <[EMAIL PROTECTED]> wrote:
>
> does anyone have protocols / method of sephadex resin. about swelling the
> resin use of buffers, elution of sample from
Look at the paper (search in pubmed) by B. M. Schick and F. Jurnak on
cryoprotection of TuTs. He changes the reservoir in 24 steps using a 24
well cryschem plate. There are some references there that are important
too.
Jackie Vitali
On Mon, Jun 23, 2008 at 8:14 PM, Ji lee <[EMAIL PROTECTED]> wro
Dear,
I have a crystal diffracted anisotrophically. I tested with a few
different cryo conditions like oil, glycerol in different
concentration to get a better data but these conditions didn't help
any.
Using capillary method improved the diffraction (isotrophic) but the
crystal couldn't survive d
dear john,
the reference for this in my opinion is still the 1994 paper by george and
wilson where they describe the use of the second virial coefficient parameter
as a metric for quantifying the potential that certain conditions have to
favour crystallisation.
see for instance bonete and
Reduce adds hydrogens to HETATMs according to its "hetdict", which is a text
file in the PDB Format for the chemical component dictionaries. A
dictionary file is available for download from the url that you cited.
Reduce will find the hetdict from either it's compiled path location, the
location
Would like to know what experiences people have had using in line
refractive index and multiangle light scattering detectors for membrane
protein purification. Specifically, if the information gathered from
such experiments has been useful in optimizing solution conditions for
membrane protein prep
On Mon, Jun 23, 2008 at 4:09 AM, Exec <[EMAIL PROTECTED]> wrote:
> Dear All,
>
> I have econo column from Biorad and AKTA FPLC system. how shall i connect
> the econo column to AKTA FPLC system i.e what are the different
> connections / tubings to be used since akta system the tubing are all
> sma
You can coexpress the enzyme with chaperone (e.g GroEL and GroES)very
often this is very successful. (There are bunches of ways though...)
Best wishes,
Arundhati
2008/6/23 yangliuqing <[EMAIL PROTECTED]>:
> Hello,everyone!
> Sorry to bother you!
> Now I have an enzyme cloned in p
You should add some salt when you anneal!!!
The duplex is highly negatively charged, so adding even a
small amount (like 10mM NaCl) will help with charge
screening, thus making the two strands less repellant to
each other. Buffer is also always a good idea. At low pH
and high temp you'll hy
Hello,everyone!
Sorry to bother you!
Now I have an enzyme cloned in pQE2 ,expressed in E.Coli M15 at room
temperature,but there are a lot of inclusion bodies,perhaps 90%,it is a big
trouble because I need to get much enzyme.
Is there anyone knows how to solve?
Thank you very
The primary reason they don't have much on the Sephadex resins is
that the are quite old-fashioned (and also relatively cheaper than
most of their high-end resins). However, old Pharmacia/LKB had some
really great manuals on chromatography that are still quite useful
for new biochemists.
Perhaps I should have mentioned that I'm working with an MTZ file
created from merged scalepack input... I was simply wondering I can't
seem to use sortmtz to perform the k->h, h->k, -l->l transformation and
have it merge these directly? It doesn't seem to want to merge from a
merged file...
Roni
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Any basic chromatography (for proteins) text will have those. It's
generally not hard - if you have dry sephadex just resuspend it in 10-20%
alcohol, stir well (but gently!) and let sit overnight, then pack the
column, wash, and go.
Artem
> does anyone have protocols / method of sephadex resin. a
What you need is a tubing and connector kit. Every decent lab has one, and
you also usually can order these in part or whole from FPLC/HPLC
manufacturers. The two types of female fittings are then attached to a
short length of tubing and presto.
Artem
> Dear All,
>
> I have econo column from Bior
GE sells all sorts of adaptors. I use those designed to let you connect
syringes to the ÄKTA. Never had problems running Econo columns.
Check out page 676/677 of your GE catalog (2006 edition) under the
heading 'tubing fittings and unions'.
Andreas
Exec wrote:
Dear All,
I have econo c
does anyone have protocols / method of sephadex resin. about swelling the
resin use of buffers, elution of sample from the resin. i have referred to
the GE healthcare website, it gives details of other resins well, but
sephadex resin not much detail is given.
kindly guide
Dear All,
I have econo column from Biorad and AKTA FPLC system. how shall i connect
the econo column to AKTA FPLC system i.e what are the different
connections / tubings to be used since akta system the tubing are all
small
sized and econo column tubing are large capacity.
does anyone have an
Partha Chakrabarti wrote:
Hi,
Is it possible to cut out a spherical map around a heavy atom site
directly without having to make bones or building something? I want to
place that in a larger P1 cell and use that for phased molecular
replacement (Phaser / Molrep).. can someone point me how to do
Roni Gordon wrote:
I have used the CCP4 program "re-index" to add a screw axis before (e.g.
P6 -> P61, P2 -> P21), but never to change the point group (e.g. PG4 ->
PG422).
I initially re-indexing the reflections without error, but refmac5 later
complained that there were too many reflections for
Sampath Natarajan wrote:
Dear All,
I am refining a structure with 2.5A resolution by refmac5. I
could find the solution by MR using molrep. After fitting the model, I
refined the structure again with 0.3 weighting term, but the output PDB file
shows many splits in the residues. So I us
I''ve got a structure with a pile of sulphate ions in the PDB file,
all in chain X. However, in the molecule selection window, these
sulphates do not appear as "monomers", nor does chain X appear in the
chain list: should they not be there?
I can select them explicitly as "//X/*", but I'm p
Dear Alexandra,
we are using the VELP FOC incubators
(http://www.velp.com/en/product/5/Environment_line/101/Refrigerated_incubators/143/FOC_225I/)
around 2000 Euros.
Demetres
Alexandra Deaconescu wrote:
Hello everyone:
I know this question has been posted before, but what models of
crysta
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