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

            王賢純 , jian Zhou a , b , , Jixian Xiong a , Jianglin Li a , Sha Huang a , Hai Zhang a , Quanze He a , Yong Lin a , Ping Chen a , Xianchun Wang a , * , Songping Liang a

            Analytrical Biochemistry 404(2010)204-210,-0001,():



            A gel absorption-based sample preparation method for shotgun analysis of membrance protcome has been developed. In this new method, membrance proteins solbilized in a strating buffer containing a high con-centration of sodium dodecyl sulfate(SDS) were directly entrapped and immobilized into gel. After the detergent and other salts were removed by washing the proteins were subjected to in-gel digestion and the tryptic peptieds were extracted and analyzed by capilary liquid chromatorgraphy 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 only avoided the protein loss and the adverse protein modifications during gel embedment but also improved the subsequent in-gel digestion and the recoery of trypic pepides, particularly the hydropho-bic peptieds, thereby facilitating the identification of membrane proteins, especially the integral mem-brane proteins compard with the conventiona tube-ge; digestion method the newl developed method increased the mumbers of identified membrane proteins and integral membrane proteins by 25.0% and 30.2%, respectively demonstration that the method is of broa practicability in gel-based shot-gun analysis of membrane proteome.


            【免責聲明】以下全部內容由[王賢純]上傳于[2010年10月08日 17時47分12秒],版權歸原創者所有。本文僅代表作者本人觀點,與本網站無關。本網站對文中陳述、觀點判斷保持中立,不對所包含內容的準確性、可靠性或完整性提供任何明示或暗示的保證。請讀者僅作參考,并請自行承擔全部責任。


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