Schuck P, Millar D B
Molecular Interactions Resource, BEPS, OD, National Institutes of Health, Bethesda, Maryland 20892, USA.
Anal Biochem. 1998 May 15;259(1):48-53. doi: 10.1006/abio.1998.2638.
A new method is described that allows measurement of the molar mass of the solute within 15 to 30 min after start of a conventional long-column sedimentation equilibrium experiment. A series of scans of the concentration distribution in close vicinity of the meniscus, taken in rapid succession after the start of the centrifuge run, is analyzed by direct fitting using the Lamm equation and the Svedberg equation. In case of a single solute, this analysis of the initial depletion at the meniscus reveals its buoyant molar mass and sedimentation coefficient with an accuracy of approximately 10% and provides gross information about sample heterogeneity. This method can be used to study macromolecules that do not possess the prolonged stability needed in conventional sedimentation equilibrium experiments and it can increase the efficiency of sedimentation equilibrium experiments of previously uncharacterized samples.
本文描述了一种新方法,该方法可在传统长柱沉降平衡实验开始后的15至30分钟内测量溶质的摩尔质量。在离心机运行开始后,对弯月面附近的浓度分布进行一系列快速连续扫描,并使用Lamm方程和Svedberg方程通过直接拟合进行分析。对于单一溶质,对弯月面处初始耗尽的这种分析可揭示其浮力摩尔质量和沉降系数,准确度约为10%,并提供有关样品异质性的总体信息。该方法可用于研究在传统沉降平衡实验中不需要长时间稳定性的大分子,并且可以提高对以前未表征样品进行沉降平衡实验的效率。