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Updated: Jun 3, 2026

Hybrid Clear/Blue Native Electrophoresis for the Separation and Analysis of Mitochondrial Respiratory Chain Supercomplexes
Published on: May 19, 2019
Hybrid Isoelectric Focusing Using Mixed Synthetic-Carrier Ampholyte-Immobilized pH Gradient Gels
Abstract:
An exciting recent innovation in isoelectric focusing (IEF) has been the development of Immobiline reagents (LKB) (1) for the preparation of polyacrylamide gels containing immobilized pH gradients (IPGs). The Immobilines are a series of seven acrylamide derivatives with the general structure: [Formula: see text] where R contains either a carboxyl (acidic Immobilines) or a tertiary amino (basic Immobilines) group, forming a series of buffers with different pK values (3.6, 4.4, 4.6, 6.2, 7.0, 8.5, 9.3). IPGIEF gels are prepared in the same manner as conventional gradient polyacrylamide gels using a concentration gradient, stabilized with a density gradient (e.g., glycerol), of the Immobiline reagents mixed with acrylamide and N,N'-methylene bisacrylamide (Bis) under the control of a suitable gradient forming device. Thus, during polymerization the buffering groups forming the pH gradient are covalently attached and immobilized via vinyl bonds to the backbone of the polyacrylamide matrix.
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