Today we discussed a new kind of electrophoresis: Polyacrylamide Gel Electrophoresis (PAGE) is a process that uses the same principle of agarose gel electrophoresis, but it uses a polyacrylamide gel, a thinner, more expensive kind of gel that provides a higher resolution than its agarose counterpart.
Specifically we were supposed to use it to run protein samples and in two ways.
- A native gel, in which the proteins migrate at different rates depending on their size (molecular weight), tertiary structure, and charge.
- A denaturing gel, in which the proteins are denatured with high temperature and kept denatured by SDS contained in the electrophoresis buffer, so their rate of migration through the gel depends exclusively on size.
The goal was to estimate the size of the green fluorescent protein (GFP) by comparing its migration through each gel with the migration of a molecular weight ruler (a "protein ladder") loaded onto the same gel. Unfortunately we didn't have the reagent to stain the gels (coomasie), so we didn't actually run the gels, although we did talk about the process.
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