Poster: Emerging technologies

Abs # 913: Robust sample preparation for the proteomic analysis of multiple plant species and tissues

Presenter:	Juehne, Thomas , Tjuehne@sial.com
Authors	Juehne, Thomas  (A)   Song, Keming  (A)
Affiliations:	(A): Sigma-Aldrich

Sample preparation is a key step in proteomic research. The combination of highly resolving separation techniques coupled with mass determination of polypeptides and bioinformatics has provided the basic inroads to specific proteomes. Proteomic research in plants presents many unique challenges. Plants, due to phenolics, polyphenolics, tannins, waxes and other interfering substances, require special consideration with respect to extraction of biological molecules. Extraction of proteins usually requires pre-treatment or protein precipitation for efficient extraction and removal of interfering substances. Developed is a method for the fast, simple and efficient extraction of proteins from plant providing total protein or fractionation of hydrophilic and hydrophobic protein pools. Extraction of proteins based on sequential stringency solubilizations with chaotropic, reducing and denaturing reagents. Highly improved solubilzation reagents provide increased levels of extraction of protein from multiple plant tissues and species. The protocol does not require any ultra centrifugation or aqueous polymer two-phase partitioning (APTP). Extraction performance provides protein pools both hydrophilic and highly hydrophobic deeply lipid bound proteins. Sample produced are compatible with downstream proteomic analysis such as two-dimensional electrophoresis and mass spectroscopy.