Schafer-Nielsen C, Svendsen P J, Rose C
J Biochem Biophys Methods. 1980 Aug;3(2):97-128. doi: 10.1016/0165-022x(80)90032-9.
The isotachophoresis principle provides unique opportunities for rational designs of fractionation procedures involving molecules. Theoretically any two charged molecules that are soluble under the experimental conditions involved can be physically separated if their electrophoretic net mobilities differ only slightly in the electrophoresis medium used. A theoretical and practical outline is presented that enables the reader to set up this fractionation system and on a rational basis develop fractionationprocedures for a given set of charged macromolecules by isotachophoresis with simple and well characterized ampholytes as spacer substances. The planning of preparative experiments in this approach is based on results obtained from rapid analytical screens on a microgram scale. The report includes an appendix containing the theoretical basis for computation of buffer compositions in the isotachophoretic steady state with mono/polyvalent constituents in systems involving one or more counterions and controlled amounts of interfering ions.
等速电泳原理为涉及分子的分级分离程序的合理设计提供了独特的机会。理论上，在相关实验条件下可溶的任何两种带电分子，如果它们在所用的电泳介质中的电泳净迁移率仅略有不同，就可以进行物理分离。本文给出了一个理论和实践大纲，使读者能够建立这个分级分离系统，并在合理的基础上，通过使用简单且特性明确的两性电解质作为间隔物质的等速电泳，为给定的一组带电大分子制定分级分离程序。这种方法中制备实验的规划基于微克级快速分析筛选获得的结果。该报告包括一个附录，其中包含在涉及一种或多种抗衡离子和可控量干扰离子的系统中，计算等速电泳稳态下含有单价/多价成分的缓冲液组成的理论基础。
