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Hi Ramasubbu,
i would suggest you to try co-purification. Often the protein complex
are cloned together its always good but its time consuming. You can
also try to mix up the pelletes before sonicating. It works fairly good
for me. You just need to take one litter pellet of each protein, mix it
in the same buffer and break the cells.
Good luck
regards
Rajesh
N. Ramasubbu wrote:
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Hi all:
I am trying to crystallize a complex of two proteins by mixing equal
concentration of the two.
At equimolar ratio or at 10mg/mL, for example, I get a precipitate
right away after mixing.
These two form a tight complex and even in other experiments using gel
filtration we have seen precipiatation
and when run on a gel, we see two bands with appropriate molecular
masses.
Molecular weight of one is around 56 kDa and the other one is 20kDa.
I would like to know of methods that one could try to keep the complex
in solution so that it can be crystallized.
Both are highly soluble proteins and can be crystallized individually
with out any difficulty.
Both form crystals using MPD at about 40%. Any help would be realy
appreciated.
Thanks a lot
subbu
