title: 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.
authors.affiliation: Jerry Lewis Muscle Research Centre, Royal Postgraduate Medical School, UK.
authors.forename: M J
authors.initials: MJ
authors.lastname: Dunn
authors.forename: K
authors.initials: K
authors.lastname: Patel