alexa The Optimized Conditions of Two Dimensional Polyacrylam
ISSN: 0974-276X

Journal of Proteomics & Bioinformatics
Open Access

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Research Article

The Optimized Conditions of Two Dimensional Polyacrylamide Gel Electrophoresis for Serum Proteomics

Fengming Gong, Shufang Liang*, Chunmei He, Guobo Shen, Yuhuan Xu, Zhizhong Xu, Zhenling Wang, Xiaohua Wu, Yuquan Wei

These authors contribute equally to this work

State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, West China Medical School, Sichuan University, Chengdu, 610041, The People’s Republic of China.

*Corresponding Author:
Dr. Shufang Liang.
State Key Laboratory of Biotherapy
West China Hospital, Sichuan University
1# Keyuan Road 4, Gaopeng Street
Chengdu 610041, China
Fax : +86 28 85164060,
E-mail : [email protected]

Received Date: July 14, 2008; Accepted Date: August 06, 2008; Published Date: August 13, 2008

Citation: Fengming G, Shufang L, Chunmei H, Guobo S, Yuhuan X, et al. (2008) The Optimized Conditions of Two Dimensional Polyacrylamide Gel Electrophoresis for Serum Proteomics. J Proteomics Bioinform 1:250-257. doi:10.4172/jpb.1000032

Copyright: © 2008 Fengming G, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

 

Abstract

Serum proteome analysis provides a potential promising approach in disease diagnosis and therapeutic monitoring. However, the large dynamic range of proteins, high-abundant proteins, excess of salt and lipid in serum makes the analysis very challenging. Therefore, it is imperative to improve the dissolution of proteins in twodimensional gel electrophoresis (2-DE), and enhance the ability to analyze the proteome of serum under a wide variety of physiological conditions. This study examined the effects of various combinations of depletion highabundant protein, precipitated means, hot SDS treatment and IPG strips with different pH on 2-DE map for mouse serum. Finally the removal of high-abundant proteins in serum by the ProteoExtractTM Albumin Removal column, ethanol precipitation, the heating with 2.5% SDS and 2.3% DTT to denature sample at 95oC for 3 min, and IEF on pH 4-7 IPG strips (17cm) with 100 ?g depleted serum proteins are generally recommended for serum proteome analysis on 2-DE by silver staining, which can effectively improve the resolution and intensity of low-abundant proteins. The optimized conditions help to produce a better reference 2-DE gel of serum samples for following identification potential novel disease markers.

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