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双向电泳分析鱼类组织。

DIGE Analysis of Fish Tissues.

机构信息

Department of Gametes and Embryo Biology, Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, Olsztyn, Poland.

出版信息

Methods Mol Biol. 2023;2596:303-322. doi: 10.1007/978-1-0716-2831-7_21.

Abstract

Two-dimensional difference gel electrophoresis (2D-DIGE) appears to be especially useful in quantitative approaches, allowing the co-separation of proteins of control samples and proteins of treated/disease samples on the same gel, eliminating gel-to-gel variability. The principle of 2D-DIGE is to label proteins prior to isoelectric focusing and use three spectrally resolvable fluorescent dyes, allowing the independent labeling of control and experimental samples. This procedure makes it possible to reduce the number of gels in an experiment, allowing the accurate and reproducible quantification of multiple samples. 2D-DIGE has been found to be an excellent methodical tool in several areas of fish research, including environmental pollution and toxicology, the mechanisms of development and disorders, reproduction, nutrition, evolution, and ecology.

摘要

二维差异凝胶电泳(2D-DIGE)似乎特别适用于定量方法,可以在同一张凝胶上同时分离对照样品和处理/疾病样品的蛋白质,消除凝胶间的可变性。2D-DIGE 的原理是在等电聚焦之前对蛋白质进行标记,并使用三种光谱可分辨的荧光染料,从而可以独立标记对照和实验样品。该程序可以减少实验中的凝胶数量,实现多个样品的准确和可重复的定量。2D-DIGE 已被发现是鱼类研究的多个领域中的一种极好的方法学工具,包括环境污染和毒理学、发育和障碍机制、繁殖、营养、进化和生态学。

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