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Electrophoretic Separation of Proteins

Published: June 12, 2008 doi: 10.3791/758


In this video, we demonstrate a method for electrophoretic separation of proteins using poly-acrylimide gel electrophoresis (PAGE).


Electrophoresis is used to separate complex mixtures of proteins (e.g., from cells, subcellular fractions, column fractions, or immunoprecipitates), to investigate subunit compositions, and to verify homogeneity of protein samples. It can also serve to purify proteins for use in further applications. In polyacrylamide gel electrophoresis, proteins migrate in response to an electrical field through pores in a polyacrylamide gel matrix; pore size decreases with increasing acrylamide concentration. The combination of pore size and protein charge, size, and shape determines the migration rate of the protein. In this unit, the standard Laemmli method is described for discontinuous gel electrophoresis under denaturing conditions, i.e., in the presence of sodium dodecyl sulfate (SDS).


The complete text protocol for this experimental approach is available in Current Protocols in Molecular Biology.

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The authors have nothing to disclose.


Electrophoretic Separation Proteins Protein Separation Electrophoresis Complex Mixtures Subunit Compositions Protein Samples Purification Of Proteins Polyacrylamide Gel Electrophoresis Electrical Field Pore Size Acrylamide Concentration Protein Charge Protein Size Protein Shape Migration Rate Laemmli Method Discontinuous Gel Electrophoresis Denaturing Conditions Sodium Dodecyl Sulfate (SDS)
Electrophoretic Separation of Proteins
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Cite this Article

Chakavarti, B., Chakavarti, D.More

Chakavarti, B., Chakavarti, D. Electrophoretic Separation of Proteins. J. Vis. Exp. (16), e758, doi:10.3791/758 (2008).

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