Hi Eswar
Firstly, I would certainly try crystal seeding into random screens if
you haven't already tried it. Refs below.
Secondly, it's very convenient to grow the crystals under oil, and to
soak the organic solvents into the drops, through the oil. This makes
it much easier to harvest the crys
I would definitely try gelfiltration (how do you get rid of the cleaved tag
anyway, sample buffer exchange?) but especially ion exchange. A homogeneous
sample on SDS-PAGE and/or gelfiltration is often not (at all) homogeneous in
ion exchange. Beyond that i would make some point mutations on surf
Depending on how big your broccoli is I would simply crush them and mount as
big of a piece as possible and see how well it diffracts.
I assume you have tried already:
- glycerol
- changing protein:reservoir ratios
- temperature
- adding oil to either / and the reservoir, your drop
-crushed up som
