I am doing ammonium sulphate precipitation step for partial purification of asparaginase. At different salt concentrations of 60 and 70% protein pellet didn't settle down. it floats even after centrifugation . I am not getting the pellet. visible mass is suspended in solution.
Is there a lot of glycerol or other dense substance (e.g. sucrose) in your sample? That could prevent the precipitated protein from pelleting. A solution to this problem would be to dilute the suspension with a solution containing the same concentration of ammonium sulfate, but not the dense substance, to reduce the density of the suspension.
It may help you in resolving your problem, try your asparaginase pptn at a different % of AS concentrations and maintaining the pH.
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