Protein Purification and Characterization Techniques PDF
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Uploaded by AccomplishedMemphis8323
2018
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Summary
This document describes protein purification and characterization techniques. It covers topics including homogenization and differential centrifugation. The methods described involve isolating protein from cells and organelles, eliminating contaminants, and obtaining a pure sample of the proteins.
Full Transcript
Chapter 5 Protein Purification and Characterization Techniques Extracting Cellular Protein: Rupturing Cells © 2018 Cengage Learning. All Rights...
Chapter 5 Protein Purification and Characterization Techniques Extracting Cellular Protein: Rupturing Cells © 2018 Cengage Learning. All Rights Reserved. Isolation of Proteins from Cells Many different proteins exist within one cell Many techniques used to eliminate contaminants and obtain pure sample First step – release protein from cells and organelles Homogenization used to break open cells © 2018 Cengage Learning. All Rights Reserved. Homogenization Potter-Elvehjem homogenizer Thick-walled test tube Tight-fitting plunger passed through tube Cells broken open Strained to remove extracellular material © 2018 Cengage Learning. All Rights Reserved. Differential Centrifugation Ruptured cells centrifuged several times Force of gravity increased each time Differential spin speeds allow for particle separation © 2018 Cengage Learning. All Rights Reserved. Homogenization Other techniques Grind tissue in blender with suitable buffer Using sonication - Sound waves break open cells Purification Once cellular contents released, purify protein of interest © 2018 Cengage Learning. All Rights Reserved.
