Characteristics of protein-storing cells associated with a 67 kDa protein in

A protein extraction method compatible with proteomic analysis for the euhalophyte Salicornia europaea

“…Triton X-100, a nonionic detergent that is efficient in breaking lipid-lipid and lipid-protein interactions [1,19,30], was included in the BPP buffer to isolate membrane proteins. Compared to the method described by Tian and colleagues [32,33], the concentrations of Triton X-100 and b-mercaptoethanol were increased from 0.1 and 1% to 1 and 2%, respectively. The optimized concentrations of EDTA, PVPP and sucrose were based on the Phe method described by Saravanan and Rose [8].…”

“…Ascorbic acid (vitamin C), as well as b-mercaptoethanol [5], are strong reducing agents and can inhibit the phenolic oxidation efficiently (2-DE Manual, GE Healthcare). Ascorbic acid and sodium borate, originally introduced to extract storage proteins from barks of poplar tree [38], were used to isolate proteins from the recalcitrant tissues of olive leaf [39], and vegetative storage proteins from tropical trees [32,33]. The oxidization of polyphenol to polyquinones and the activity of many enzymes were inhibited under the deoxidized condition generated by b-mercaptoethanol, vitamin C, and PVPP.…”

“…The extraction buffer used in the BPP protocol was modified based on the recipe for extracting soluble proteins from trees by Tian and co-workers [32,33], with modifications as described below. Triton X-100, a nonionic detergent that is efficient in breaking lipid-lipid and lipid-protein interactions [1,19,30], was included in the BPP buffer to isolate membrane proteins.…”

“…The protein extraction buffer was optimized from the published ones [32,33]. In short, 1 g of frozen lyophilized tissue powders was resuspended in 3 mL ice-cold extraction buffer of 100 mM Tris (pH 8.0) containing 100 mM EDTA, 50 mM borax, 50 mM vitamin C, 1% PVPP w/v, 1% Triton X-100 v/v, 2% b-mercaptoethanol v/v and 30% sucrose w/v.…”