One-dimensional electrophoresis using nondenaturing conditions.
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.
Gallagher
S R
SR
Motorola Corporation, Tempe, Arizona, USA.
D000818:Animals; D002021:Buffers; D002138:Calibration; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods* / Q000592:standards; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification*
D002021:Buffers; D011506:Proteins