Preparative 2D Gel Electrophoresis with Immobilized pH Gradients: IEF of Proteins in an IEF-Dedicated Electrophoresis Unit

Citations Over Time

Abstract

INTRODUCTION This protocol describes a method for separating proteins based on their net charge using the technique of isoelectric focusing (IEF) on immobilized pH gradient (IPG) gels, providing the first dimension of the 2D separation. In this protocol, the IPG gels are focused using self-contained instruments for IEF. These high-voltage systems allow fewer manipulations of the IPG gels, resulting in less error, strip mix-up, contamination, air contact, or urea crystallization. Because rehydration and IEF can be performed consecutively within a single unit, these two steps can be performed unattended overnight. Finally, faster separations and sharper focusing are possible due to the higher voltage available in these instruments.

Related Papers

• → Aberrant Migration of Lipopolysaccharide in Sodium Dodecyl Sulfate/Poyacrylamide Gel Electrophoresis(1983)49 cited
• → Simultaneous immunoblotting analysis with activity gel electrophoresis in a single polyacrylamide gel(2001)23 cited
• → Isoelectric focusing of proteins separated by SDS-polyacrylamide gel electrophoresis(1977)34 cited
• → Binding of lithium dodecyl sulfate to polyacrylamide gel at 4°C perturbs electrophoresis of proteins(1986)6 cited
• → Staining of proteins after SDS-polyacrylamide gel electrophoresis by ethidium bromide(1996)1 cited