Ohlendieck Kay
Department of Biology, Maynooth University, National University of Ireland, Maynooth, Co. Kildare, Ireland.
Methods Mol Biol. 2018;1664:17-24. doi: 10.1007/978-1-4939-7268-5_2.
Gel-based proteomics has been widely used for the systematic cataloging of the protein constituents of defined biofluids, purified organelles, individual cell types, heterogeneous tissues and isolated organs, as well as being applied to comparative biochemical and biomedical analyses of complex biological specimens. Of the many electrophoretic techniques used in modern biochemical approaches, large-scale protein separation by difference gel electrophoresis (DIGE) has established itself as the most powerful analytical tool in comparative proteomics. Both 2-dye and 3-dye fluorescence systems with minimal or saturation labeling are routinely used. This chapter briefly describes the technical advantages of the pre-electrophoretic fluorescent labeling technique and discusses the bioanalytical usefulness of this highly successful electrophoretic method.
基于凝胶的蛋白质组学已被广泛用于对特定生物流体、纯化细胞器、单个细胞类型、异质组织和分离器官中的蛋白质成分进行系统编目,也被应用于复杂生物样本的比较生化和生物医学分析。在现代生化方法中使用的众多电泳技术中,差异凝胶电泳(DIGE)进行大规模蛋白质分离已成为比较蛋白质组学中最强大的分析工具。常规使用具有最小或饱和标记的双染料和三染料荧光系统。本章简要描述了电泳前荧光标记技术的技术优势,并讨论了这种非常成功的电泳方法在生物分析中的实用性。