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Chapter 14

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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 gelsThis 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. SDS-PAGEChapter 14Objectives124Chapter 14 SDS-PAGE is widely used to analyze the proteins in complex extracts.

Discontinuities between the stacking and running gels underlie the resolving power of the SDS-PAGE gels The Laemmli (1970) SDS-PAGE system can be con-sidered a 3-component system. The stacking and running (resolving) gels have different pore sizes, ionic strengths and pHs. The third component is the electrophoresis buffer

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