2009-09-09_-_Lecture_3

2009-09-09_-_Lecture_3 - Biochemistry 461 FS 2009 Michigan...

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Biochemistry 461 FS 2009 Michigan State University Protein Purification September 9, 2009 Lecture 3 Protein Purification General strategy for protein purification (5 Basic Steps) 1. Rupture cells to release proteins Freeze Sonicate French press (not for coffee) Vortex with small glass beads 2. Collect the proper fraction 3. Purify protein of interest by selecting for characteristics of that protein 4. Determine purity of protein – Is protein pure? No? Repeat step 3 Yes? Continue 5. Characterize purified protein 2
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Step 2: Collect the proper fraction Protein Purification 3 • Proteins differ from each other in a number of characteristics, each of which can be used to separate the protein of interest from the complicated mixture in a tissue sample or laboratory culture: • Solubility • Charge at a given pH (pI) •S ize • Binding to a particular substance or material • In all cases you are going to have to check each fraction for your protein of interest Protein Purification Step 3: Purify the protein of interest from other proteins and cellular material
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2009-09-09_-_Lecture_3 - Biochemistry 461 FS 2009 Michigan...

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