| 抑制off-gel分离中18O标记回标的方法研究 |
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| 引用本文： | 熊嫣,邓玉林. 抑制off-gel分离中18O标记回标的方法研究[J]. 北京理工大学学报, 2015, 35(S1): 1-5 |
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| 作者姓名： | 熊嫣 邓玉林 |
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| 作者单位： | 北京理工大学 生命学院, 生物医药成分分离与分析北京市重点实验室, 北京 100081,北京理工大学 生命学院, 生物医药成分分离与分析北京市重点实验室, 北京 100081 |
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| 基金项目： | 国家重大科学仪器设备开发专项资助项目(2012YQ040140) |
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| 摘 要： | off-gel分离法和18O标记是蛋白质组学研究中常用的分离和标记方法,通过对大鼠纹状体蛋白酶解后的肽段进行off-gel分离,以及基于超滤后的18O标记与off-gel分离的结果表明,off-gel对复杂生物样品具有良好的聚焦和分离效果,超滤去除游离酶后,18O标记的肽段没有发现明显回标现象,16O与18O标记的肽段1:1混合后,90%以上的蛋白16O/18O比值为1.04±0.16,因此,对复杂样品的考察结果说明基于超滤的18O标记可以与off-gel分离联合使用,扩大了18O标记在分离方法中的选择范围,能够应用于定量蛋白质组学的研究中.
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| 关 键 词： | 18O标记 游离胶分离 蛋白质组学 超滤 |
| 收稿时间： | 2015-03-20 |
Method for Suppressing 18O Labeling Back-Exchange in off-gel Fractionation |
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| XIONG Yan and DENG Yu-lin. Method for Suppressing 18O Labeling Back-Exchange in off-gel Fractionation[J]. Journal of Beijing Institute of Technology(Natural Science Edition), 2015, 35(S1): 1-5 |
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| Authors: | XIONG Yan and DENG Yu-lin |
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| Affiliation: | School of Life Science, Beijing Institute of Technology, Beijing Key Laboratory of Bio-Separation and Bio-Analysis, Beijing 100081, China and School of Life Science, Beijing Institute of Technology, Beijing Key Laboratory of Bio-Separation and Bio-Analysis, Beijing 100081, China |
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| Abstract: | Off-gel fractionation and 18O labeling are widely used in proteomics research. In this study, two methods were applied in tryptic peptides from rat striatum protein. Results show that off-gel has better separation and isoelectric focusing for complex biological samples. Furthermore, 18O labeled peptides have no apparent back-exchange, and over 90% proteins from equal amounts of 16O and 18O labeled peptides mixture have a 16O/18O ratio of 1.04±0.16 after ultrafiltration, indicating no apparent effect on quantification and 18O labeling is compatible with off-gel separation based on ultrafiltration, which can provide an alternative separation method for 18O labeling and play important role in quantitative proteomics research. |
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| Keywords: | 18O labeling off-gel fractionation proteomics ultra-filtration |
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