I am extracted total protein from Rice plant by Phenol extraction buffer followed by overnight precipitation in ammonium acetate, pellet down the pellet, 3 times washing with methanol, 2 times washing with acetone and then air-drying of pellet. I have to dissolve this pellet of protein in 20mM ammonium bicarbonate solution, but it is not getting dissolved. I also used 1X PBS and 20mM Tris buffer but the pellet is not dissolved properly.
Anyone dissolved protein in ammonium bicarbonate solution please suggest me regarding this, if any modifications needed. I have to move further with ammonium bicarbonate solution.
Since the proteins are completely denatured by phenol extraction, I think it is unlikely that they will dissolve in a simple, fairly neutral, aqueous buffer like ammonium bicarbonate. You might be able to dissolve them in SDS-PAGE sample buffer.
Thanking you Shapiro for your critical suggestion. Ya it may be possible to dissolve it in SDS-PAGE buffer, I will try for this. But I have to proceed with ammonium bicarbonate buffer only, so suggest regarding this.
Sometimes the aggregation happen after centrifugation... To solve this problem we applied ultrasound bath then the pellet dissolv
What did you finally do to dissolve the pellet. I am facing the same problem.
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