Protein Purification and Determination (1).pdf

Protein Purification and Determination (1).pdf - Protein...

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Protein Purification and Determination of Protein Structure
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Lesson Learning Outcomes Upon completion of this lecture, students should be able to: Know the procedure of protein extractions Differentiate types of chromatography Know the electrophoresis
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Protein Purification Differential centrifugation Used for separation of cell components using different speed of centrifugation Different centrifugational force will separate larger cell components from smaller ones Low speed to separate large cell components High speed to separate smaller cell components
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Homogenization Before the real purification steps can begin, the protein must be releases from the cells and subcellular organelles through differential centrifugation In general, separation techniques focus on size, charge and polarity. Proteins will be purified either through size-exclusion chromatography, affinity chromatography, isoelectric points, High Performance Liquid Chromatography (HPLC).
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Chromatography Greek chroma = color Greek graphein = to write Early technique used to separate plant pigments with easily visible colors It is based on the fact that the different compounds can distribute themselves to different phases Stationary phase & mobile phase
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Column chromatography
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  • Spring '16

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