Dear Eric,
There are several essential variables that govern protein chromatography
(whether affinity, ion exchange, sizing, or other). It would be silly of me
to reproduce a protein chromatography handbook here, so instead I would just
list some practical pointers:
If your column is e.g. 10-15
Eric,
I second Chun's reply. As the Volumetric flowrate (ml/min) is equal
to the linear flowrate times the area of the column, and the linear
flowrate is fixed for a particular resin (Sigma is terrible at providing
this value for their resins) you basically just need to increase the
Donghui,
We have several membrane proteins crystallized in salt-based
conditions, actually in high sodium citrate. Yes, LiCl or lithium
formate are good alternatives, but also try lithium citrate, as
citrate also depresses the freezing point quite well.
Good luck,
Michael
There are many excellent review articles about cryocrystallography and
cryoprotectants. Do labs generally have these articles handy in a methods
folder? Do lab heads help their colleagues by making them read them?
Mineral oil is generally not a good oil to use because it changes volume
too
Hi Donghui,
I just want to draw your attention to the following article. It sums up
suggestions which were already made by others for using concentrated
(lithium) salt solutions for cryoprotection:
Rubinson, K.; Ladner, J.; Tordova, M. Gilliland, G. Cryosalts:
suppression of ice formation in
Hi everyone,
I received a lot of positive e-mails about my post on cryo
methods. One theme was, Well, what are the papers I should read?
Blatant self-promotion
I know of one such paper published in Methods. I have made the PDF of it
downloadable from our web site at
Have you tried the various oils ?
As a last resort Greek olive oil if paratone and others fail.
How about sugars ?
Stay away from propanol, your crystals will start floating through your
wells with very rapid movements. I once had a 30% 2-propanol crystal
condition and mounting crystals only
Dear Lokesh,
Actually I have tried to dip my crytal directly into mineral oil, no ice
ring, but also no diffraction, which might indicate crystal packing under
mineral oil is not stable. Anyway, thanks a lot for your valuable
information. Other friends mention the use of LiCl or Lithium formate
Are you familiar with pm3d? I couldn't give the solution from the top of
my head, but
http://t16web.lanl.gov/Kawano/gnuplot/plotpm3d-e.html
seems a good source of information.
--
Tim Gruene
Institut fuer anorganische Chemie
Tammannstr. 4
D-37077 Goettingen
GPG Key ID = A46BEE1A
On Wed, 16
On Wed, 16 Jan 2008, Jacob Keller wrote:
I would like to plot as a heat or color map on the 2d rectangle
[...] cannot figure out how to do this in gnuplot or anywhere,
consider gnuplot pm3d (not readily obvious for googling, is it)
http://t16web.lanl.gov/Kawano/gnuplot/plotpm3d-e.html
-bryan
On Wednesday 16 January 2008 13:27, Jacob Keller wrote:
Hello all,
It might just take you a few minutes to tell me how to do this:
I have a set of data in three columns xyz (resid1, resid2, value), which I
would like to plot as a heat or color map on the 2d rectangle formed by
the
You don't need much of a machine to be a Linux backup server. I built mine from
a salvage box (a discarded PIII machine) and a couple of EIDE HDDs. The OS went
on a 80 Gbyte drive and the data is stored on a 500 Gbyte drive. The machine is
located in another building on campus. My master server
Dear Shivesh,
Try some buffer screening.
If you have a real-time PCR machine nearby you can easily screen
96 buffer conditions in about an hour.
Check out:
Ericsson et al. Analytical Biochemistry 2006 Aug;357(2):289-98
Best regards,
Martin
On Aug 30, 2007, at 6:06 PM, shivesh kumar wrote:
Perhaps someone would barter with you for your reagents?
On Wednesday 11 July 2007 18:28, Eric Dollins wrote:
Thank you to all who replied to my original post. Here is the result.
Original query:
Sorry for the off-topic question.Does anyone know a way to just
buy the spin columns from
There are several sources.
You can often get them from the manufacturer of your miniprep kit
(the catalog # is written on the side of the bag they come in within
the kit). Many times, they don't list in their catalog that they
will sell it separately, but you can call and get them to give
Bill:
My $0.02 (speaking as someone who has not been happy lately with Apple
tech support and customer care).
This strikes me as a completely unacceptable response. Unless I'm
missing something, if the machine is no longer under warranty then you
are simply asking for a fee-for-service repair,
It's not that simple, it seems. First off, there are numerous reports
that Apple repair centers are routinely taking out third-party RAM,
sending along a form letter that that RAM was the likely culprit.
Giving the customer the option of installing RAM does not mean one
can expect to
My $0.02 (speaking as someone who has not been happy lately with Apple
tech support and customer care).
This strikes me as a completely unacceptable response. Unless I'm
missing something, if the machine is no longer under warranty then you
are simply asking for a fee-for-service repair,
We also currently have a laptop at Apple for repair. The repair center
charges a flat rate of $329, no matter what the repair entails.
We have a similar deal here, but my understanding is that if the actual
cost of parts exceeds $329, you get charged the difference. Since it was a
hard drive,
Steve,
Steve Lane wrote:
Given the current situation at Apple, particularly their shift in focus
and revenue percentage from computers to other types of devices,
i.e. iPods, either/both of the above reasons for refusal to do the work
are plausible.
are you saying one shouldn't buy one's
We have a similar deal here, but my understanding is that if the
actual cost of parts exceeds $329, you get charged the difference.
Since it was a hard drive, I didn't get to test the hypothesis. If
the repair for a 2 year old laptop is much more than that, there is
no point in doing the
Hi Bill,
This sounds very unusual. We have 3rd party RAM in almost all of our
Macs, and I've never had a repair request denied by AppleCare. This
is an allowed DIY (do it yourself) installation in most if not all
their recent machines. It's common knowledge now that hard drives
fail
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