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Protein Purification

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Biochemistry 2280A
Eric Ball

Protein PurificationNeed to isolate the protein from contaminants Take advantage of the properties to isolate them Assay methodneed a test that is specific to the proteintook a mixture of proteinsrun this test and you detect the protein of interest enzymetest for enzyme activity Start with large scale methods and work towards small scale methods Preparative methodsbreak up the materialcells are wholeproteins are inside the cell so break open the cellStart on a large scalestart with a lot of leavesorgan etc After you move through the preparative methodsmove to analytical which is a smaller scalesee how pure the protein isHomogenizebreak open Pressureisolate the protein from yeastput it under pressurecracking noise and breaks open cells To make sure its maintaining activity and structurebufferwhen you test it it will be maintaineddont want the protein to be ruined by the end Cells are broken opennext step is to centrifuge the material Spins very quickly The technique they use is differential centrifugationspin the homogenate at different speeds Different speeds because the different parts of the cell will pellet under different g forces depending on its size Smallest stuffpellet at
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