Article ID Journal Published Year Pages File Type
1184418 Food Chemistry 2015 7 Pages PDF
Abstract

•Protein sample preparation of extracted oil bodies for Tricine–SDS–PAGE.•Proteins in oil body suspension can be well solubilized by SDS.•SDS/protein mass ratio of 1.52/1 is efficient for protein solubilization.•Higher or lower SDS/protein is unbeneficial for protein solubilization.•About 99% of proteins can be recovered on Tricine–SDS–PAGE gel.

A simple and efficient method for preparing Tricine–SDS–PAGE protein sample of extracted oil bodies (OBs) was supplied: OB suspension was vortexed with SDS buffer (pH 6.8) for 2 min at room temperature with SDS/protein of 1.52/1 (w/w), which could be analyzed by Tricine–SDS–PAGE after simple treatments (dilution and 2-mercaptoethanol). At SDS/protein of 1.52/1, about 95% of proteins in soybean OB suspension were solubilized, whereas residual 5% of proteins were weakly bound to SDS-destroyed OBs; proteins in destroyed OBs might be further solubilized by SDS in the gel and cathode buffer of Tricine–SDS–PAGE, causing about 99% of proteins in soybean OB suspension recover on Tricine–SDS–PAGE gel, which was better than acetone (89%) and diethyl ether (96%) harvested protein samples. Higher or lower SDS/protein was unbeneficial for protein solubilization from OBs. Additionally, the above method was also better than organic solvent method for peanut, sesame, and rapeseed OB suspensions.

Related Topics
Physical Sciences and Engineering Chemistry Analytical Chemistry
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