Protein purification by Off-Gel electrophoresis
| Autor: | Hubert H. Girault, Joël S. Rossier, Alexandra Ros, Philippe Michel, Rosaria Ferrigno, Jan van Oostrum, Frédéric Reymond, Michel Faupel, Hervé Mees |
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| Jazyk: | angličtina |
| Rok vydání: | 2002 |
| Předmět: |
Gel electrophoresis
Two-dimensional gel electrophoresis Chromatography Proteome Isoelectric focusing Chemistry Proteins Fractionation Prefractionation Hydrogen-Ion Concentration Isoelectric purification Biochemistry 660.6 Electrophoresis Isoelectric point Bacterial Proteins Electric field Protein purification Escherichia coli Electrophoresis Gel Two-Dimensional Isoelectric Point Molecular Biology Serpins |
| DOI: | 10.1002/1615-9861(200202)2:2<151::aid-prot151>3.0.co;2-9 |
| Popis: | A novel free-flow protein purification technique based on isoelectric electrophoresis is presented, where the proteins are purified in solution without the need of carrier ampholytes. The gist of the method is to flow protein solutions under an immobilised pH gradient gel (IPG) through which an electric field is applied perpendicular to the direction of the flow. Due to the buffering capacity of the IPG gel, proteins with an isoelectric point (pI) close to pH of the gel in contact with the flow chamber stay in solution because they are neutral and therefore not extracted by the electric field. Other proteins will be charged when approaching the IPG gel and are extracted into the gel by the electric field. Both a demonstration experiment with pI markers and a simulation of the electric field distribution are presented to highlight the principle of the system. In addition, an isoelectric fractionation of an Escherichia coli extract is shown to illustrate the possible applications. |
| Databáze: | OpenAIRE |
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