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| Establishment of Two-dimensional Electrophoresis System of Brassica oleracea L. var. capitata L. Stigma Protein |
| ZENG Jing1，CHEN Song1，ZHU Li-quan1*，GAO Qi-guo2*，WANG Xiao-jia2，YANG Xiao-hong2 |
| 1College of Agronomy and Biotechnology，Southwest University，Plant Physiology and Biochemistry Laboratory，Southwest University，Chongqing 400716，China|2College of Horticulture and Landscape Architecture，Southwest University，Key Laboratory of Horticulture Science for Southern Mountainous Regions，Ministry of Education，Chongqing 400715，China |
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Abstract A two-dimensional electrophoresis（2-DE）has been widely applied in proteomics research．The purpose of this study was to establish a suitable 2-DE system for proteomic analysis of Brassica oleracea L. var. capitata L. stigma．The stigmas being harvested one day before flowering was used in the experiment．We optimized the parameters of 2-DE including extraction methods，IPG strip，sample loading，isoelectric focusing conditions and SDS-PAGE gel concentration．The optimized system included the following steps：extracting the total proteins from Brassica oleracea stigma by TCA/acetone precipitation，separating the proteins with length 17 cm and pH 3-10 NL IPG strips，loading proteins samples of 150 μg followed by isoelectric focusing programⅠ，and selecting SDS-PAGE gel concentration at 12%．The 2-D images with good repeatability and high protein spot resolution were obtained．
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Received: 02 December 2011
Published: 15 February 2012
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