6 - Lecture 6: Electrophoretic and centrifugal separations...

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Unformatted text preview: Lecture 6: Electrophoretic and centrifugal separations of proteins Tuesday, August 17, 2010 Movement of charged molecules in an electrical field Like chromatography, electrophoresis is a fundamentally important technique in protein research. Electrophoresis uses an electric field to separate proteins based upon their charge and size: Remember from physics that: F elec = qE where the Force (F) on an ion is determined by the charge q on the ion and the electrical field strength, E. Tuesday, August 17, 2010 Also, F frict = vf a frictional force (F frict ) opposes the movement on the ion, dependent upon the velocity, v, and a frictional coefficient, f. In a constant field, the two forces balance each other, such that qE = vf Tuesday, August 17, 2010 Thus, an electrophoretic mobility can be described as: μ = v/E = q/f IN SHORT, the migration of an ion (a protein) through an electric field is dependent upon its charge (q) and and shape (f). Tuesday, August 17, 2010 Currently, in laboratories, proteins are visualized using a technique called GEL ELECTROPHORESIS. While there are different types of gels available (for instance, DNA is often run through a gel composed of a polysaccharide called agarose), proteins are most often run in POLYACRYLAMIDE gels: The polymerization of acrylamide and bisacrylamide yields a gel containing many continuous pores ....
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This note was uploaded on 11/12/2010 for the course CHEN 3320 at Colorado.

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6 - Lecture 6: Electrophoretic and centrifugal separations...

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