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[ccp4bb]: Co-crystallisation with hydrophobic ligands
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Many thanks to everyone who emailed, suggestions were really helpful.
Here is a summary of responses:
1. Co-concentrate ligand with protein: (This is actually something I
tried before posting my question; it seems to work with some ligands
but with others concentration causes both protein and ligand to
Gitay Kryger & Jeroen Brandsen:
2. Soak/Co-crystallise the crystal in drop containing solid ligand.
3. If Kd is slow, then excess of ligand is not required and unbound
ligand will be at low effective concentration anyway. (In fact adding
excess will not give much greater occupancy).
4. Use 1.3 dimethyl -2-imidazolidinone (DMI) instead of DMSO/DMF etc.
DMI is miscible in water and can tolerate up to 15%(v/v) against
protein. (I hadnt heard of this solvent before, but im going to try
5. Stir the ligand extensively in buffer or buffer+DMSO, then spin down
and use the solution in co-xtallization.