One‐Dimensional Electrophoresis Using Nondenaturing Conditions
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Open Access
- 1 July 1999
- journal article
- unit
- Published by Wiley in Current Protocols in Molecular Biology
- Vol. 47 (1) , 10.2B.1-10.2B.11
Abstract
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.Keywords
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