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Related Experiment Videos

Finite element simulation of Off-Gel trade mark buffering.

Isabelle L Arnaud1, Jacques Josserand, Joël S Rossier

  • 1Laboratoire d'Electrochimie, Département de Chimie, Ecole Polytechnique Fédérale de Lausanne, Lausanne, Switzerland.

Electrophoresis
|October 10, 2002
PubMed
Summary

A minimum Immobiline concentration is essential for maintaining pH stability in gels during Off-Gel electrophoresis. This finding is crucial for understanding proton and ampholyte distribution in buffered systems.

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Area of Science:

  • Electrochemistry
  • Biophysical Chemistry
  • Separation Science

Background:

  • Ampholytes are crucial in electrophoresis for separating proteins.
  • Immobilized pH gradients (IPGs) are used in electrophoresis to stabilize pH.
  • Understanding protonation and diffusion is key to optimizing electrophoretic separations.

Purpose of the Study:

  • To investigate the pH evolution between an ampholyte solution and an Immobiline gel.
  • To determine the influence of Immobiline concentration on proton and ampholyte distribution.
  • To establish a theoretical framework for Off-Gel electrophoresis.

Main Methods:

  • Finite element simulation in an iterative scheme.
  • Formulation of a ten-species model including transient diffusion and equilibrium kinetics.

Related Experiment Videos

  • Modeling protonation of aqueous ampholyte solutions adjacent to immobilized acid moieties.

    • Main Results:

    • A minimum Immobiline concentration of 10(-2) M is necessary for pH stability in closed systems.
    • This minimum concentration is required when the isoelectric point difference of ampholytes is 4 and initial ampholyte concentration is in the micromolar range.
    • The study illustrates pH changes and distribution of protons and ampholytes.

    Conclusions:

    • Finite element simulation provides a theoretical basis for Off-Gel electrophoresis.
    • Immobiline concentration is a critical factor in achieving stable pH gradients for electrophoresis.
    • The findings guide the optimization of Off-Gel electrophoresis protocols.