Thanks you for the suggestions, Patrick and Matthias. I was actually
wondering if any of the components from the seeding solution actually were
important but your explanations sound more logical.
I apologize for the large attachment. I did not realize that it was so big.
Have a nice weekend !
Ma
Mahesh, this is a very interesting and slightly controversial question.
One approach is to mix together all of the crystals that you have in the
initial screen. The idea at the beginning of the project is to get as many
diverse hits as possible - you can worry about crystal size, space group
and
Dear Mahesh,
cross-seeding into other conditions often works but still the chances
are much lower than seeding the same form.
Best wishes,
Matthias
(Please desist from sending so large attachments to half the planet!)
-
Dr. Matthias Zebisch
Division of
Does anyone have a TTP Mosquito robot Deck Configuration file for XQ-P-96S-C
plates (3 wells x 96) made by MiTeGen?
--
David M. Mueller
Biochemistry and Molecular Biology
The Chicago Medical School
Rosalind Franklin University of Medicine and Science
Green Bay Road
North Chicago, IL 60064
Two 3-Year Postdoctoral Scientist Positions
National Institute for Medical Research (NIMR), London
NIMR will become part of the Francis Crick Institute (www.crick.ac.uk)
Informal enquires may be made by email:
Peter Rosenthal
pros...@nimr.mrc.ac.uk
https://cryoem.nimr.mrc.ac.uk/
(1) Cryomicr
Cedric,
I suspect you are correct in that the 2 mM ATP, which will have
significant absorption at 280 nm, is contributing a high background
absorbance. The Bradford method is notoriously idiosyncratic, as it
depends on hydrophobic content of the protein, and is affected by
substances that cha
The solution might be as simple as measuring absorbance at 290 nm. We routinely
measure concentrations in the presence of 100 uM ATP ( usually without
concentrating though). That said 2 mM ATP is a lot more than we usually deal
with and may interfere even at 290 nm. The interference from ATP dr
Dear CCP4 Users,
An update for the CCP4-6.4.0 series has just been released, consisting of the
following changes:
* pointless
* fixed crash when using reference data with no alternative indexing
* with multiple files, include columns present in any file
* added crash mes
Try preparing your Bradford standard curve with your protein buffer
(including the ATP). This is how I reliably get around detergent and other
compounds that interfere.
Hope this helps,
Kelly
***
Kelly Daughtry, Ph.D.
IRTA Fellow
Mechanisms of
Dear Cedric,
Try the BCA protein assay. We get reliable results with this, consistent with
amino acid analysis. I should say that this is using the 2ml format. We do
not get reliable results with the microplate method. We use 5 μl samples and
our regular buffer is not too different from the
Dear all,
We're doing some crystallization trials on a protein that requires 2mM
ATP in the buffer and we are having trouble measuring
reliably/reproducibly protein concentration.
This is a real problem for optimization (last screen failed because of
excessive protein concentration compared to
On inspecting the code, it turns out that this is not much a bug as a feature.
I will try to make it an ex-feature, but it will take a little work
The work-around is to convert the .sca file to MTZ first, using scalepack2mtz.
Note that a merged Scalepack file (unlike an unmerged one) does contai
12 matches
Mail list logo
