| 双向电泳分离技术的比较和优化 |
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| 引用本文： | 舒宏,康晓楠,李山,刘银坤,秦雪.双向电泳分离技术的比较和优化[J].广西医科大学学报,2009,26(1). |
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| 作者姓名： | 舒宏 康晓楠 李山 刘银坤 秦雪 |
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| 作者单位： | 1. 广西医科大学第一附属医院临床医学实验部,广西医科大学检验系,南宁,530021
2. 复旦大学中山医院肝癌研究所,生物医学研究院癌症研究中心,上海,200032 |
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| 基金项目： | 国家高技术研究发展计划(863计划)，国家自然科学基金，广西高校优秀人才资助计划，广西自然科学基金，广西医疗卫生重点课题 |
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| 摘 要： | 目的:比较优化人血清蛋白双向电泳(2-DE)分离技术,提高2-DE对血清蛋白的分辨能力.方法:收集健康人群标本,分别进行多次双向电泳,比较高丰度蛋白的去除前后及传统的考马斯亮蓝染色和SYPRO-ruby荧光染色两种不同的染色方法等因素对双向电泳图谱质量的影响.结果:考马斯亮蓝染色和SYPRO-ruby荧光染色相比,血清2-DE图谱检测到平均蛋白质点数分别是(324±25)(n=3)和(785±30)(n=3);去除高丰度蛋白前后比较,未做处理的血清、去除蛋白和IgG血清的2-DE图谱检测到平均蛋白点数分别是(501±25)(n=3)和(813±22)(n=3).结论:去除高丰度蛋白并结合SYPRO-ruby荧光染色可以大大提高血清低丰度蛋白的解析能力.此方法优化了血清蛋白质2-DE技术,为进一步开展疾病血清蛋白质组研究奠定了基础.
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| 关 键 词： | 双向电泳 血清蛋白质组 去除高丰度蛋白 染色 |
COMPARISON AND OPTIMIZATION OF TWO-DIMENSIONAL POLYACRYLAMIDE GEL ELECTROPHORESIS(2-DE) TECHNIQUE |
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| Shu Hong,Kang Xiaonan,Li Shan,et al..COMPARISON AND OPTIMIZATION OF TWO-DIMENSIONAL POLYACRYLAMIDE GEL ELECTROPHORESIS(2-DE) TECHNIQUE[J].Journal of Guangxi Medical University,2009,26(1). |
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| Authors: | Shu Hong Kang Xiaonan Li Shan |
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| Institution: | Shu Hong,Kang Xiaonan,Li Shan,et al.(Central Laboratory,the First Affiliated Hospital of Guangxi Medical University,Nanning 530021 China) |
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| Abstract: | Objective:To compare and optimize a two-dimensional polyacrylamide gel electrophoresis(2-DE) technique,and to enhance the resolution for serum proteomics.Methods:Serum samples were obtained from healthy donors. The effectiveness of different conditions was compared repeatedly according to the final results on 2-DE maps. Depleting high abundance protein,Coomassie Brilliant Blue and SYPRO ruby fluorescent dye staining on the quality of 2-DE maps were compared.Result:The average spots in the Coomassie Brillian... |
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| Keywords: | Two-dimensional polyacrylamide gel Electrophoresis (2-DE) Serum proteomics depletion high-abundance protein staining |
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