On Wed, 2011-11-16 at 09:26 +, Tom Murray-Rust wrote:
> That way you should be able to
> quickly identify any hits that are due to salt, and which are likely
> to be your protein.
Just a footnote to Tom's excellent comment:
It is possible to have actual protein crystals to grow alongside sal
A thing we frequently forget is that phosphate can be a precipitating agent
try a phosphate grid screen, just like you would with ammonium sulfate.
If your protein likes PBS, it may want to crystallize with phosphate
See Enrico Stura's footprint screen for example
On Tue, Nov 15, 2011 at 5:25 PM,
Of course PBS should not be a first choice for screning crystal.
But I will try all kinds of buffer until I got the structure.
Nobady can tell you that you can't get crystal in the PBS buffer. and I will
shot everything I have if I have enough beam time.
Good luck.
Hi JK,
As mentioned, phosphate salts are the main disadvantage - you can get
round this by setting up two drops per well: one with your protein in
PBS, and the other with PBS only. That way you should be able to
quickly identify any hits that are due to salt, and which are likely
to be your protei
Jayakrishnan Nandakumar
Sent: Wednesday, November 16, 2011 12:26 AM
To: CCP4BB@JISCMAIL.AC.UK
Subject: [ccp4bb] Crystallizing protein sitting in PBS
Hi All,
I have an RNA-binding protein that I can purify out of bacteria
in PBS (Phosphate
Hi All,
I have an RNA-binding protein that I can purify out of bacteria in PBS
(Phosphate buffered saline;137 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 2 mM
KH2PO4), but which is insoluble in Tris/NaCl-based buffers. My guess would
be that the inorganic phosphates (by mimicking RNA) are binding the prote
