Podcast
Questions and Answers
What tool is used to fill the column?
What tool is used to fill the column?
- Pipette
- Micropipette
- Microscope
- Syringe (correct)
What is the purpose of filling the column?
What is the purpose of filling the column?
- To sterilize the column
- To fill the column with buffer to just above the bed support screen (correct)
- To wash the column
- To equilibrate the column
Where is the GF buffer injected into?
Where is the GF buffer injected into?
- Column outlet tubing (correct)
- Column body
- Column inlet tubing
- Column support screen
What is the final level of buffer in the column?
What is the final level of buffer in the column?
What is filled with GF buffer?
What is filled with GF buffer?
What is the primary factor in selecting a gel filtration matrix?
What is the primary factor in selecting a gel filtration matrix?
What is the main purpose of gel filtration?
What is the main purpose of gel filtration?
What is the goal of selecting a gel filtration matrix?
What is the goal of selecting a gel filtration matrix?
What is gel filtration used for?
What is gel filtration used for?
What is the basis of separation in gel filtration?
What is the basis of separation in gel filtration?
What is the minimum molecular weight of proteins that are completely excluded from Sephadex G-75?
What is the minimum molecular weight of proteins that are completely excluded from Sephadex G-75?
What is the term for the volume of buffer required to elute a given substance?
What is the term for the volume of buffer required to elute a given substance?
Which of the following is a characteristic of Sephadex G-75?
Which of the following is a characteristic of Sephadex G-75?
What happens to proteins with molecular weights greater than 70,000 when they are passed through Sephadex G-75?
What happens to proteins with molecular weights greater than 70,000 when they are passed through Sephadex G-75?
What is the purpose of measuring the elution volume of a compound?
What is the purpose of measuring the elution volume of a compound?
What is the purpose of allowing the elution to proceed at an appropriate hydrostatic pressure or flow rate?
What is the purpose of allowing the elution to proceed at an appropriate hydrostatic pressure or flow rate?
What is the purpose of collecting the column effluent?
What is the purpose of collecting the column effluent?
What is the benefit of using an automated fraction collector?
What is the benefit of using an automated fraction collector?
What is the role of the column effluent in the separation process?
What is the role of the column effluent in the separation process?
What is the relationship between the hydrostatic pressure or flow rate and the separation process?
What is the relationship between the hydrostatic pressure or flow rate and the separation process?
What happens to Cyt09rorne c during the process?
What happens to Cyt09rorne c during the process?
Which of the following proteins elute in order of decreasing molecular weight?
Which of the following proteins elute in order of decreasing molecular weight?
What is true about the elution of the other proteins?
What is true about the elution of the other proteins?
Why does Cyt09rorne c elute last?
Why does Cyt09rorne c elute last?
What is the relationship between the molecular weight of the proteins and their elution order?
What is the relationship between the molecular weight of the proteins and their elution order?
Flashcards
Gel Filtration
Gel Filtration
A technique used to separate molecules based on size.
Gel Filtration Matrix
Gel Filtration Matrix
A material used in gel filtration to separate molecules.
Molecular Size
Molecular Size
The key factor determining separation in gel filtration.
Elution Volume
Elution Volume
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Sephadex G-75
Sephadex G-75
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Column Preparation
Column Preparation
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Buffer
Buffer
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Exclusion Limit
Exclusion Limit
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Column Effluent
Column Effluent
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Fraction Collector
Fraction Collector
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Hydrostatic Pressure
Hydrostatic Pressure
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Flow Rate
Flow Rate
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Protein Separation
Protein Separation
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Elution Order
Elution Order
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Molecular Weight
Molecular Weight
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GF Buffer
GF Buffer
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Column Outlet Tubing
Column Outlet Tubing
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Bed Support Screen
Bed Support Screen
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Buffer Level
Buffer Level
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Study Notes
Gel Filtration (GF) Gel Matrix
- The choice of matrix depends on the range of size of molecules to be separated and the goal of the separation.
- Filling the column with GF buffer is done using a syringe, injecting until the column is filled to just above the bed support screen.
Characteristics of Sephadex G-75
- Proteins with molecular weights greater than 70,000 are completely excluded.
- Volume of buffer required to elute a substance is known as the elution volume, Ve.
Elution Process
- Elution proceeds at an appropriate hydrostatic pressure or flow rate.
- Column effluent is collected using an automated fraction collector.
- Elution order: proteins are partially included and elute in order of decreasing molecular weight, with the smallest (Cytochrome c) eluting last.
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Description
Learn about the principles of gel filtration, including the choice of matrix, and its applications in separating molecules based on size. Understand the importance of gel filtration in various scientific fields.
