I have the previous model with NVIDIA GeForce 9400M and it's been just fine -
on the desktop I use a Zalman stereo and it's great.
According to Pierce TCEP is "more" tolerant of nickel and cobalt. However, TCEP
is inactivated by other metals, namely copper, magnesium, silver and zinc.
Dan
Daniel A. Bonsor,
Boston Biomedical Research Institute,
64 Grove Street,
Watertown,
MA 02472 USA
That's a possibility. Do you know if the same coloration would arise if there
is TCEP instead of DTT or b-ME? I usually have TCEP as the reducing agent.
Thanks Fillip.
Cheers,
Shiva
--- On Fri, 9/24/10, Filip Van Petegem wrote:
From: Filip Van Petegem
Subject: Re: [ccp4bb] protein turns br
>
> I also have a similar observation for proteins purified by Ni-NTA column.
> After concentrating the sample eluted from the Ni-NTA column, I see a
> brownish-yellowish tinge closer to the bottom of the filter with colorless
> buffer on top. This is observed even for non-metal binding and non-FeS
Hi All,
I also have a similar observation for proteins purified by Ni-NTA column. After
concentrating the sample eluted from the Ni-NTA column, I see a
brownish-yellowish tinge closer to the bottom of the filter with colorless
buffer on top. This is observed even for non-metal binding and non-F
and possibly the opposite to reduction which is oxidation - do you have cys
residues? Perhaps your DTT or TCEP got exhausted? Remember you can add up to
10mM (not the traditional token 1mM). But remember to neutralize TCEP. Other
possibility is adventitious metals as these give strong charge tra
Yellow or brown tinge is very common especially if you have used sonication to
lyse the cells. It helps to wash the cells with buffer or even high salt and
spin them down again before lysis. If you want brown then you should see what
happens when you work with yeast.
Probably flavins or some
It could be caused by iron contamination in one of your buffers. We used to
buy glycerol in a metal canister and metal would leach into the glycerol.
Because of this, one protein that I worked with would turn yellow, even at
relatively low concentrations. I did not have this issue when using
glyc
[FeS] clluster ?
Or some metal bound to your protein ?
Is it dark brown or yellowish brown ? What's your protein concentration ? > 25
mg/ml ?
Jürgen
-
Jürgen Bosch
Johns Hopkins Bloomberg School of Public Health
Department of Biochemistry & Molecular Biology
Johns Hopkins Malaria Research Instit
Maybe you should give us a hint about the identity of your protein (if you
dare;-)). I'm sure there are folks around who may be able to say whether or
not your protein is supposed to be brown. You can't expect too much help if you
don't provide (m)any details.
Cheers, Bert
On 9/24/10 4:34
Hi Sandy,
If your protein is soluble (i.e. we're not dealing with brown
precipitate), it is possible that the brown color is due to a bound
iron-sulfur cluster. In this case, your protein was always brown, but
you couldn't see the color when it was very dilute. As a quick test,
you could t
Hi Sandy,
Is there any possibility of your protein to contain an iron-sulfur cluster?
Best
Ricardo
sometime it does happen becoz of protein aggregation, or reducing environment.
but it may be your protein color as well that visible during concentration
Vikrant
From: sandeep
To: CCP4BB@JISCMAIL.AC.UK
Sent: Fri, 24 September, 2010 2:04:54 PM
Subje
Dear all,
I have purified protein from E.coli. expression system. the protein has been
purified with three independant columns. Now during concentration step using
amicon, the protein shows brown colour. what could be the reason.
best regards and Thanks,
sandy
For the sake of a thought experiment we can assume the refinement
is implemented with infinite-precision arithmetic. And for actually
...
terms in the 2n'd derivative matrix are ignored?). Two ncs-related atoms
can move in agreement with the ncs-restraint without penalty from the
near-infinite n
Dear All,
We're pleased to announce that registration for this coming January's
CCP4 Study Weekend meeting is now open. The meeting will take place from
January 5th to 7th, 2011 at University of Warwick in the UK. The topic
of the meeting will be "Model building and Refinement and Validation".
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