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Gel absorption-based sample preparation for the analysis of membrane proteome by mass spectrometry

Gel absorption-based sample preparation for the analysis of membrane proteome by mass spectrometry,10.1016/j.ab.2010.05.013,Analytical Biochemistry,Ji

Gel absorption-based sample preparation for the analysis of membrane proteome by mass spectrometry   (Citations: 1)
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A gel absorption-based sample preparation method for shotgun analysis of membrane proteome has been developed. In this new method, membrane proteins solubilized in a starting buffer containing a high concentration of sodium dodecyl sulfate (SDS) were directly entrapped and immobilized into gel matrix when the membrane protein solution was absorbed by the vacuum-dried polyacrylamide gel. After the detergent and other salts were removed by washing, the proteins were subjected to in-gel digestion and the tryptic peptides were extracted and analyzed by capillary liquid chromatography coupled with tandem mass spectrometry (CapLC–MS/MS). The results showed that the newly developed method not only avoided the protein loss and the adverse protein modifications during gel embedment but also improved the subsequent in-gel digestion and the recovery of tryptic peptides, particularly the hydrophobic peptides, thereby facilitating the identification of membrane proteins, especially the integral membrane proteins. Compared with the conventional tube-gel digestion method, the newly developed method increased the numbers of identified membrane proteins and integral membrane proteins by 25.0% and 30.2%, respectively, demonstrating that the method is of broad practicability in gel-based shotgun analysis of membrane proteome.
Journal: Analytical Biochemistry - ANAL BIOCHEM , vol. 404, no. 2, pp. 204-210, 2010
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