Lab 4 - Polyacrylamide Gel Electrophoresis (PAGE) Under...

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Polyacrylamide Gel Electrophoresis (PAGE) Under Denaturing Conditions 1
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Polyacrylamide Gel Electrophoresis (PAGE) Under Denaturing Conditions University of Texas at Austin Department of Chemistry and Biochemistry Introduction Gel electrophoresis allows charged molecules to migrate through a solution once a current is placed going through the gel. The way the molecules migrate down the gel, due to charge density, can be used to separate and determine molecules. The gels used for electrophoresis can vary by pore size, allowing the user to construct the experiment to their standards. The gels can also vary by type; starch, agarose, and polyacrylamide are common types of gels. We used a polyacrylamide gel because they are great for proteins and can achieve very high resolution. The addition of a sample loading buffer to each of the samples allowing the denaturation of the proteins to occur; it prevents disulfide bond formation and prevents multi- subunit complexes. 4 Once the proteins have run on the gel, typically in the vertical direction, the use of a staining solution is used to help visualize the protein movement in the gel. The use of each
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This note was uploaded on 09/07/2009 for the course CH N/a taught by Professor Genemcdonald during the Spring '09 term at University of Texas at Austin.

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Lab 4 - Polyacrylamide Gel Electrophoresis (PAGE) Under...

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