Increase in Local Protein Concentration by Field-Inversion Gel Electrophoresis

Methods Mol Biol. 2019:1855:211-227. doi: 10.1007/978-1-4939-8793-1_19.

Abstract

Proteins that migrate through cross-linked polyacrylamide gels (PAGs) under the influence of a constant electric field experience negative factors, such as diffusion and nonspecific trapping in the gel matrix. These negative factors reduce protein concentrations within a defined gel volume with increasing migration distance and, therefore, decrease protein recovery efficiency. Here, we describe the enhancement of protein separation efficiency for up to twofold in conventional one-dimensional PAG electrophoresis (1D PAGE), two-dimensional (2D) PAGE, and native PAGE by implementing pulses of inverted electric field during gel electrophoresis.

Keywords: Field-inversion gel electrophoresis; Forward pulse time; Pulsed-field gel electrophoresis; Reverse pulse time; Separation efficiency; Two-dimensional polyacrylamide gel electrophoresis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cross-Linking Reagents
  • Electrophoresis, Gel, Pulsed-Field / methods*
  • Molecular Weight
  • Proteins / isolation & purification*

Substances

  • Cross-Linking Reagents
  • Proteins