At our last synchrotron trip, the beamline staff suggested that the problem
was due to moisture accumulation in the dry shipper. They recommended
storing them inverted (for a few weeks, if I recall), and/or putting a
supply of dry air in the dewer. Haven't tried it yet!
Nat
On Thu, Jul 11,
I can't resist bashing 10.8 a little. I don't know if I would rush to
upgrade... I find the removal of 'save as' to be very annoying
(holding down option brings it back). Also the way Text edit would
hide the scroll bar by default was annoying when looking through log
files (easily changed
Here is an incubator about the size of the Ecotherms but much cheaper:
http://www.tritechresearch.com/DT2-MP-38.html
We have one for insect cell culture that has worked great for several
years, and there is no vibration. However for crystallization we have
BOD incubators from Fisher.
Nat
On
I mentioned to to Chris already, but we use nothing but HyClone
SFX-Insect powder. We make 20-30 L batches, sterilize with a large
peristaltic pump and a disposable Millipak filter from Millipore. We
never have contamination problems that are due to preparing our own
media from powder. We buy
I use a 10 ml GE-healthcare Ni-sepharose FF column, packed myself. I
strip it with EDTA after a run and then recharge before the next one.
It's has probably been used 30+ times, and I have no plans or
repacking it anytime soon, it still works as good as it ever did. We
purify secreted proteins
Hi,
It can, just do
fm-mode
select rmsd
I am curious though, I have heard that it is 'better' to build in
units of absolute density, but I couldn't find any values. Does any
one have a suggestion as to what absolute electron density setting is
'correct' for an Fo-Fc difference map? Or do you
I use ImageJ for this purpose. There is a flag for something like
'use virtual stacks' or 'use virtual memory' that I needed to prevent
crashing.
Nat
On Mon, Mar 7, 2011 at 5:30 PM, Sean Seaver s...@p212121.com wrote:
Dear Mark,
I put together a post about creating a movie using PyMOL,
We use the Millipore Spiral-wound TFF1 and TFF2 cartridges for
concentrating 10-20L of insect cell media. They work great, don't
cost all that much money, and you don't need the expensive holder they
sell. You can use a ring stand and clamp the tubing directly to the
cartridge. Our cartridges
We don't have a problem getting them to stick to the plates in
serum-free media, or in 5% FBS media. The more challenging part is
getting the plating density just right, too low and the plaques are
too big, to high and they are too small. Or if the cells dry out, or
if your agarose overlay is
.
Jay
-Original Message-
From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of
Nathaniel Clark
Sent: Wednesday, March 30, 2011 11:38 PM
To: CCP4BB@JISCMAIL.AC.UK
Subject: Re: [ccp4bb] titering baculovirus ?
We don't have a problem getting them to stick to the plates
chitra.shin...@gmail.com wrote:
Hi,
We have observed this virus going off within 2 months as well. ...any idea
why this happens?
Chitra
On 31 March 2011 18:07, Nathaniel Clark nathanielcl...@gmail.com wrote:
We do adherent if we have a small volume of low titer virus, but as
soon as we have
be no need to do any viral amplification.
Cheers,
Chun
Accelagen
-Original Message-
From: Nathaniel Clark nathanielcl...@gmail.com
Sender: CCP4 bulletin board CCP4BB@JISCMAIL.AC.UK
Date: Thu, 31 Mar 2011 13:20:14
To: CCP4BB@JISCMAIL.AC.UK
Reply-To: Nathaniel Clark
First, I don't think glycosylation occurs on lysine residues.
You should try to crystallize it as is, if that doesn't work, try to
remove the glycosylation with some or all of the following: EndoH,
PNGaseF, EndoF1, EndoF3. We have used them all to various degrees of
efficiency to remove glycans
Bei,
How do you concentrate your media? We use tangential flow filtration.
If you get a good filter (Millipore Spiral wound TFF is one example)
it goes pretty quick, in ~2 hours you should be able to process
4L(concentrate+ dilute several time in buffer). We secrete proteins
from insect cells,
Hi,
I have soaked various crystals at that pH with various sugars and
never had a problem with oxidation of the sugar. I wouldn't worry
about it. What group do you think will be oxidized?
Nat
On Fri, Mar 12, 2010 at 5:35 AM, Paul Lindblom
lindblom.p...@googlemail.com wrote:
Hi,
I am trying
I have wondered if placing a layer of oil over the drop would help
solve the problem of the crystals moving around. Haven't tried it,
but don't people harvest from a microbatch tray by dragging the loop
and crystal through oil?
Nat
On Fri, Apr 9, 2010 at 11:21 AM, James Holton jmhol...@lbl.gov
MiTeGen has a set of soft plastic microtools that are inteded for this
kind of stuff, and I have had suscess using them to break out single
crystals from clumps or fused crystals. You can either load in a
pencil or mount in a base.Also nylon loops. You might need to
have one tool in each
I just tried that protocol, and had essentially all of my protein
crashed out. Any tips on optimizing the methylation reaction to
reduce precipitation? Perhaps reducing the formaldehyde or
dimethylaminoborane, shortening the incubation times, etc.?
Thanks,
Nat
On Wed, Apr 14, 2010 at 6:00 PM,
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