Hi,
To add to this thread, there are a few more easy things to try –
Try doing matrix microseeding and try doing limited proteolysis. Even though
the following link describes how we do this in our laboratory, (the C3 Facility
in Melbourne) the pages give a quick overview to both matrix microsee
Dear Amala,
You have already received a lot of good advices from expert
crystallographers. Apart from the notables, i would still read about the
protein. Is this an enzyme? If yes, what is the ligand or a co-factor? The
protein conformation would be different in apo form versus bound form.
Often,
Due the resolution of your images, it is a little bit difficult to identify
crystallization under these conditions. However, F3, F6 and F5 are
crystals, but the condition needs to be optimized. Likely, you may try
different concentration of NaCl, instead of MgCl2 and KBr, respectively.
Good luck.
Hi Katherine,
One possibility could be that you have occupancy of 0.5 and that ligand
binding at one site in the crystal distorts a second site so it cannot bind
ligand.
Occupancy refinement is not always very stable.
If you are looking at two structures on top of one another - refinement
Dear Amala,
The first thing I would try is to run your protein sequence on this server:
http://xtalpred.godziklab.org/XtalPred-cgi/xtal.pl. This is a webserver that
will compare you aa sequence to other previously crystallized proteins. The
result will also suggest which parts of your protein s
Dear Amala,
There are some good suggestions coming your way from here. I would also
recommend checking out the High-Throughput Crystallization Screening Center at
the Hauptman-Woodward Medical Research Institute or other similar services.
They run most of the common types of screens in an effi
Hi Amala,
In addition to Limbini´s suggestions.
1. You can always do MALS, SLS, Native Page or cross linking experiments to
check whether your protein is monodisperse in the solution or not. This is
very important step.
2. When you set up crystallization and if you have 40-50% drops
precipit
Dear Amala,
1. Hampton provides a precrystallisation screening kit which can be used to
determine the protein concentration to be used for crystallisation.
2. Also if you observe the screening crystallisation plate, according to
what I follow, at least there should be minimum 60 % of conditions w
Dear Prof. Rupp,
Sadly it was a comment meant seriously. A friend of mine who works in the field
of quantum mechanics had brought this article to my attention before. He has
also contacted the authors on social media where they are waging a campaign
against the use of quantum “supremacy”, which
