Podcast
Questions and Answers
What is the first step in isolating proteins from cells?
What is the first step in isolating proteins from cells?
Release protein from cells and organelles
What technique is used to break open cells?
What technique is used to break open cells?
Homogenization
What is the purpose of straining the homogenate?
What is the purpose of straining the homogenate?
Remove extracellular material
What is the purpose of differential centrifugation?
What is the purpose of differential centrifugation?
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Which of the following is NOT a technique used to homogenize cells?
Which of the following is NOT a technique used to homogenize cells?
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The force of gravity used in differential centrifugation remains constant throughout the process.
The force of gravity used in differential centrifugation remains constant throughout the process.
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What is the name of the technique used to separate particles based on their size and density?
What is the name of the technique used to separate particles based on their size and density?
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What is the ultimate goal of protein purification?
What is the ultimate goal of protein purification?
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Study Notes
Protein Purification and Characterization Techniques
- Cells contain various proteins
- Techniques isolate pure protein samples
- First step: release proteins from cells and organelles
- Homogenization breaks open cells
Homogenization Methods
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Potter-Elvehjem homogenizer:
- Thick-walled test tube
- Plunger passed through tube
- Breaks open cells
- Removes extracellular material by straining
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Blender:
- Grind tissue with suitable buffer
-
Sonication:
- Sound waves break open cells
Differential Centrifugation
- Ruptured cells centrifuged repeatedly
- Increasing force of gravity each time
- Differential spin speeds separate particles based on size and density
- Different pellets are formed at different stages of centrifugation
- Homogenate spun at low speed, and supernatant is collected.
- Subsequent supernatants are spun at increasingly higher speeds to separate progressively smaller components
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Description
This quiz covers the essential techniques for protein purification and characterization, including homogenization methods and differential centrifugation. Learn about various methods such as the Potter-Elvehjem homogenizer, blenders, and sonication, along with the principles behind differential centrifugation for isolating protein samples.
