UNIT 10.2B One-Dimensional Electrophoresis Using Nondenaturing Conditions
Published Online: 1 MAY 2001
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Molecular Biology
How to Cite
Gallagher, S. R. 2001. One-Dimensional Electrophoresis Using Nondenaturing Conditions. Current Protocols in Molecular Biology. 47:II:10.2B:10.2B.1–10.2B.11.
- Published Online: 1 MAY 2001
- Published Print: JUL 1999
Nondenaturing or “native” electrophoresis (i.e., electrophoresis in the absence of denaturants such as detergents and urea) is an often-overlooked technique for determining the native size, subunit structure, and optimal separation of a protein. Because mobility depends on the size, shape, and intrinsic charge of the protein, nondenaturing electrophoresis provides a set of separation parameters distinctly different from mainly size-dependent denaturing sodium dodecyl sulfate-polyacrylamide gel electrophoresis and charge-dependent isoelectric focusing. Two protocols are presented in this unit. Continuous PAGE is highly flexible, permitting cationic and anionic electrophoresis over a full range of pH. The discontinuous procedure is limited to proteins negatively charged at neutral pH but provides high resolution for accurate size calibration.