14: SDS-PAGE

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This lab will introduce you to SDS-PAGE, a simple and inexpensive method for resolving proteins in complex mixtures.
SDS-PAGE gels provide the starting materials for western blots and for some proteomic techniques. In this lab, you will
use SDS- PAGE to analyze the protein extracts that you prepared from yeast strains overexpressing Met and LacZ fusion
proteins.

Objectives

At the end of this lab, students will be able to:

discuss the principles that govern protein separation on discontinuous SDS- PAGE gels.
cast and run SDS-PAGE gels.
analyze the pattern of bands on a stained SDS-PAGE gel
estimate the molecular weight of a protein from its migration on SDS-PAGE gels

This lab will introduce you to SDS-PAGE (sodium dodecyl sulfate - polyacrylamide gel electrophoresis), a simple and inexpensive method for resolving proteins in complex mixtures. You will use SDS-PAGE gels to analyze the yeast protein extracts that you prepared in the last lab. Each team will make two gels. One gel will be stained with Simply Blue to visualize all the pro- teins in the extracts. The second gel will be used for western blots (Chapter 15) that will specifi- cally detect Met and LacZ fusion proteins in the extracts.