Olson Bradley J S C, Markwell John
Michigan State University, East Lansing, Michigan, USA.
Curr Protoc Protein Sci. 2007 May;Chapter 3:Unit 3.4. doi: 10.1002/0471140864.ps0304s48.
Biochemical analysis of proteins relies on accurate quantitation of protein concentration. This unit describes how to perform commonly used protein assays, e.g., Lowry, Bradford, BCA, and UV spectroscopic protein assays. The primary focus of the unit is assay selection, emphasizing sample and buffer compatibility. Protein assay standard curves and data processing fundamentals are discussed in detail. This unit also details high-throughput adaptations of the commonly used protein assays, and also contains a protocol for BCA assay of total protein in SDS-PAGE sample buffer that is used for equal loading of SDS-PAGE gels, which is reliable, inexpensive, and quick.
蛋白质的生化分析依赖于蛋白质浓度的准确定量。本单元介绍了如何进行常用的蛋白质测定，例如洛瑞法、考马斯亮蓝法、BCA法和紫外光谱蛋白质测定法。本单元的主要重点是测定方法的选择，强调样品和缓冲液的兼容性。详细讨论了蛋白质测定标准曲线和数据处理的基本原理。本单元还详细介绍了常用蛋白质测定方法的高通量改进方法，并且包含一个用于SDS-PAGE样品缓冲液中总蛋白BCA测定的方案，该方案用于SDS-PAGE凝胶的等量加样，可靠、廉价且快速。
