Determination of molecular size by size-exclusion chromatography (gel filtration).
Determination of molecular size by size-exclusion chromatography (gel filtration).
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DOI：
10.1002/0471143030.cb0505s06
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发表时间：
2001-05-01
影响因子：
--
通讯作者：
Irvine, G B
中科院分区：
文献类型：
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作者：
Irvine, G B
尺寸排阻或凝胶过滤色谱法是用于确定蛋白质尺寸的最流行的方法之一。将溶液中的蛋白质或其他大分子应用于具有确定支持介质的柱。蛋白质的行为取决于其大小和介质中孔的大小。如果蛋白质相对于孔径较小，则它将分配到介质中，并在较大的蛋白质之后从柱中出现。除了蛋白质的大小，这项技术还可用于蛋白质纯化，纯度分析和蛋白质之间相互作用的研究。在本单元中，提供了尺寸排阻高效液相色谱法（SE-HPLC）和常规凝胶过滤的方案，包括使用蛋白质标准品校准色谱柱（根据斯托克斯半径）。
Size-exclusion or gel filtration chromatography is one of the most popular methods for determining the sizes of proteins. Proteins in solution, or other macromolecules, are applied to a column with a defined support medium. The behavior of the protein depends on its size and that of the pores in the medium. If the protein is small relative to the pore size, it will partition into the medium and emerge from the column after larger proteins. Besides a protein's size, this technique can also be used for protein purification, analysis of purity, and study of interactions between proteins. In this unit protocols are provided for size-exclusion high-performance liquid chromatography (SE-HPLC) and for conventional gel filtration, including calibration of columns (in terms of the Stokes radius) using protein standards.