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Precipitating plant proteins
I am working with rhizome proteins i extract using a phenol based method. The proteins have given me good SDS PAGE bands. My problem now is that my proteins of late seem to be insoluble in urea lysis buffer (7M urea, 2M thiourea, 2% chaps) after hours of vortexing. Sometimes the proteins dissolve and i keep the solutin at -20C. Upon thawing, the proteins tend to precipita. I was planning to go to IEF and 2D electrophoresis, but with the problems i have highlighted, i am very worried. Please help me.
Re: Precipitating plant proteins
Don't let the sample stand for hrs.You should load the sample within 1hr after dissolving in buffer.
Try 10% TCA in Acetone precipitation method.
And after precipitation add rehydration buffer and let it be so for 10 min then dissolve sample slowly with an spatula.
|plant , precipitating , proteins|
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