大小排阻色谱法
折射法
多角度光散射
化学
光散射
化学计量学
散射
色谱法
折射计
激光器
分析化学(期刊)
动态光散射
光学
折射率
材料科学
物理
纳米技术
生物化学
物理化学
纳米颗粒
酶
出处
期刊:Humana Press eBooks
[Humana Press]
日期:2004-04-02
卷期号:: 113-118
被引量:13
标识
DOI:10.1385/1-59259-762-9:113
摘要
The stoichiometry of a protein complex can be calculated from an accurate measurement of the complex’s molecular weight. Multiangle laser light scattering in combination with size-exclusion chromatography and interferometric refractometry provides a powerful means for determining the molecular weights of proteins and protein complexes. In contrast to conventional size-exclusion chromatography and analytical centrifugation, measurements do not rely on the use of molecular weight standards and are not affected by the shape of the proteins. The technique is based on the direct relationship between the amount of light scattered by a protein in solution and the product of its concentration and molecular weight. A typical experimental configuration includes a size-exclusion column to fractionate the sample, a light-scattering detector to measure scattered light, and an interferometric refractometer to measure protein concentration. The determination of the molecular weight of an anthrax toxin complex will be examined to illustrate how multiangle laser light scattering can be used to determine the stoichiometry of protein complexes.
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