I was concentrating my recombinant protein with the amicon spin filters, it was completely dissolved in PBS to begin with, that was around 8mls, but I wanted it concentrated to 10 mg/ml for crystallization studies. It precipitated out of solution when I brought it down to 1 ml. Will it redissolve now if I increase the volume of PBS? Will it be still be usable for crystallization studies?
Increasing the volume will not help most likely (assuming to have irreversible aggregates). The fact that your protein precipitates out while concentrating means that you will have to restrict the concentration limit. Crystallization is a concentration process itself and you should definitely start with a lower concentration where the protein is nicely soluble and happy. Try 5 mg/ml or 2.5 mg/ml if you think the protein craps out at 10 mg/ml. Good luck !
thanks for your suggestion Sarkhel, Konig and Lesbats. I will start fresh and keep the dilution higher.
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