Fast and sensitive coomassie staining in quantitative proteomics.
Fast and sensitive coomassie staining in quantitative proteomics.
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DOI：
10.1007/978-1-61779-885-6_4
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发表时间：
2012-01-01
期刊：
影响因子：
--
通讯作者：
Metzger, Sabine
中科院分区：
文献类型：
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作者：
Dyballa, Nadine;Metzger, Sabine
通过二维凝胶电泳分离的蛋白质可以通过使用不同染色方法的凝胶内检测来可视化。理想情况下，染料应遵循线性响应曲线与蛋白质非共价结合。由于生物系统中的蛋白质浓度可以变化六个或更多个数量级（Corthals GL等人，电泳21（6）：1104-1115，2000），染色应允许检测非常低的蛋白质量。同时，必须避免饱和效应，因为它们会妨碍标准化定量。大多数蛋白质组学实验室使用考马斯、银或荧光染色。利用考马斯亮蓝染料的胶体性质，可同时实现较低纳克水平的检测限。因此，胶体考马斯染色具有易于使用、成本低和与下游表征方法（如质谱法）兼容等特点，非常适合定量蛋白质组学中的凝胶内检测方法。
Proteins separated by two-dimensional gel electrophoresis can be visualized by in-gel detection using -different staining methods. Ideally, the dye should bind non-covalently to the protein following a linear response curve. Since protein concentrations in biological systems may vary by six or more orders of magnitude (Corthals GL et al., Electrophoresis 21(6):1104-1115, 2000), the staining should allow for a detection of very low protein amounts. At the same time, saturation effects have to be avoided because they impede normalized quantification.Most proteomics laboratories apply Coomassie, silver, or fluorescent stains. Using the colloidal properties of Coomassie dyes, detection limits at the lower nanogram level can meanwhile be achieved. Characteristics like ease of use, low cost, and compatibility with downstream characterization methods such as mass spectrometry, therefore, make colloidal Coomassie staining well suited for the in-gel detection method in quantitative proteomics.