Dear all,
Thank you for the prompt and valuable suggestions. I had made a typo in my
earlier mail. I use 50mM NaCl, not 5mM NaCl. I am facing this problem only
while using DDM in buffers. My other runs, even with 8M urea run perfectly
fine.
I will try increasing the salt concentartion and degas b
If your buffer can't go through a 0.2 micron filter easily, you
shouldn't run it through your FPLC. Detergent purity may be an issue.
I also experienced problems filtering a buffer containing CHAPS from
vendor X. When I switched to Anatrace CHAPS, no more clogging.
Ho
Ho Leung Ng
University
Hi Sam,
Your version of XSCALE is OK since it prints out the string "XSCALE" (the date
is OK - my fault!).
Rp.i.m. is indeed not listed in XSCALE.LP.
Best,
Kay
--
Diese Nachricht wurde von meinem Android-Mobiltelefon mit K-9 Mail gesendet.
Thanks for the replies Harry, Joe and Kay,
@Harry
(1) I've had a look at the com files and the the run with the file from XSCALE
reaches the ROGUEPLOT stage (command line) and says mtz MERGED (Script from
file), when I press continue from that stage it switches from running to
failed. The run
Hi Nazia,
The DDM concentration you mention seems fine to me. I routinely run the
FLPC at 4C with buffers containing 0.05% (1mM) DDM and 2% glycerol without
issues. I have also used buffers with 5% glycerol in the past but I usually
adjust the flow rate, as needed and when I know all else is good
On Fri, 26 Jul 2013 15:19:57 +0100, Joe.Lyons wrote:
>
>Hi Sam,
>
>Assuming the XDS_ASCII.HKL is from XSCALE. I generally run it through
>Pointless then aimless with (a scales constant command) using something like
>the following.
>pointless -copy xdsin XDS_ASCII.HKL hklout sorted.mtz << eof >
Hi Sam,
one possibility: there was a little bug in a recent build of XSCALE that
resulted in one of the header lines of the output file to not specify the name
XSCALE - it wrote:
!GENERATED BY: July 4, 2012
and it should have been
!GENERATED BY: XSCALE (VERSION March 30, 2013)
The lack of the
Hi Sam,
Assuming the XDS_ASCII.HKL is from XSCALE. I generally run it through Pointless
then aimless with (a scales constant command) using something like the
following.
pointless -copy xdsin XDS_ASCII.HKL hklout sorted.mtz << eof > pointless.log
eof
aimless hklin sorted.mtz hklout Scaled_1.mt
The first thing is that you'll have to increase the salt concentration in your
buffer. 5 mM is way too low and may cause non-specific binding of the protein
to the resin. 100 mM is the minimum you should use. There is nothing in your
buffer that will precipitate, so you should't have to worry ab
We are trying to purify a membrane protein using different detergents (DDM,
OG etc.). We have tried using 1mM DDM in 20mM Tris, 5mM NaCl and 5%
glycerol buffer to purify the protein. however, we are facing problems in
running the buffer in 16/60 Superdex 200 pg gel filytration coloumn using
AKTA Ex
Hi Everyone,
I've been trying to process some data using Xia2, XSCALE, Pointless and
Aimless. I have a large number of datasets (over 100) so have used Xia2 to
produce the XDS_ASCII.HKL files using the 3daii pipeline. I now want to process
them manually (with a script) using XSCALE and pointles
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