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Questions and Answers
What is the purpose of protein purification?
What is the purpose of protein purification?
- To increase the protein concentration
- To enhance protein extraction efficiency
- To isolate a pure preparation of a protein for property determination (correct)
- To expand the variety of proteins within a sample
Which technique is NOT used in the preliminary steps of protein purification?
Which technique is NOT used in the preliminary steps of protein purification?
- Ultracentrifugation to separate sub-cellular organelles
- Treating with detergents like Triton X-100
- Salting out with ammonium sulfate
- Gel filtration chromatography (correct)
What is an essential factor impacting the speed of elution in protein chromatography?
What is an essential factor impacting the speed of elution in protein chromatography?
- The size of the column packing particles (correct)
- The pH of the elution buffer
- The source of the protein (tissue or microbial cells)
- The number of freeze-thaw cycles during protein extraction
Which approach is commonly employed to break open cells during protein extraction?
Which approach is commonly employed to break open cells during protein extraction?
In protein purification by ion exchange chromatography, which peptide would elute first from a cation-exchange resin?
In protein purification by ion exchange chromatography, which peptide would elute first from a cation-exchange resin?
Which factor plays a crucial role in determining the elution order of peptides in ion exchange chromatography?
Which factor plays a crucial role in determining the elution order of peptides in ion exchange chromatography?
How does the presence of Glu and Asp residues in a peptide affect its elution behaviour in ion exchange chromatography using cation-exchange resin?
How does the presence of Glu and Asp residues in a peptide affect its elution behaviour in ion exchange chromatography using cation-exchange resin?
What is the primary purpose of chromatography?
What is the primary purpose of chromatography?
Which type of chromatography involves separating molecules based on their size?
Which type of chromatography involves separating molecules based on their size?
What was the original focus of chromatography when it was first recorded in the mid-1800s?
What was the original focus of chromatography when it was first recorded in the mid-1800s?
Which field uses chromatography for athlete testing, including horses?
Which field uses chromatography for athlete testing, including horses?
What is the main principle behind affinity chromatography?
What is the main principle behind affinity chromatography?
In affinity chromatography, how are proteins that do not bind to the ligand treated?
In affinity chromatography, how are proteins that do not bind to the ligand treated?
What is used to elute bound proteins in affinity chromatography?
What is used to elute bound proteins in affinity chromatography?
What is the effect of a free ligand in affinity chromatography?
What is the effect of a free ligand in affinity chromatography?
Which type of chromatography is primarily based on bead migration rather than binding affinity?
Which type of chromatography is primarily based on bead migration rather than binding affinity?
What is the main mechanism by which proteins are separated in size exclusion chromatography?
What is the main mechanism by which proteins are separated in size exclusion chromatography?
Salting out in protein purification involves increasing solubility of proteins by adding salts.
Salting out in protein purification involves increasing solubility of proteins by adding salts.
Proteins vary in size, charge, and binding properties due to genetic engineering.
Proteins vary in size, charge, and binding properties due to genetic engineering.
The purpose of protein purification is only to determine the properties of proteins, not their activity.
The purpose of protein purification is only to determine the properties of proteins, not their activity.
Gas Chromatography (GC) is not a common method of chromatography.
Gas Chromatography (GC) is not a common method of chromatography.
High-Performance Liquid Chromatography (HPLC) is a type of column chromatography.
High-Performance Liquid Chromatography (HPLC) is a type of column chromatography.
Thin-Layer Chromatography (TLC) does not involve a stationary phase.
Thin-Layer Chromatography (TLC) does not involve a stationary phase.
Size Exclusion Chromatography is not a separation principle used in chromatographic purification.
Size Exclusion Chromatography is not a separation principle used in chromatographic purification.
In column chromatography, the length of the column does not impact the column efficiency.
In column chromatography, the length of the column does not impact the column efficiency.
The elution order of peptides in ion exchange chromatography is primarily determined by their boiling points.
The elution order of peptides in ion exchange chromatography is primarily determined by their boiling points.
Recombinant proteins are not used for Western Blot techniques.
Recombinant proteins are not used for Western Blot techniques.
Recombinant proteins are never used in animal models to identify therapeutic targets.
Recombinant proteins are never used in animal models to identify therapeutic targets.
One of the disadvantages of recombinant proteins is their lack of post-translational modifications.
One of the disadvantages of recombinant proteins is their lack of post-translational modifications.
In hydrophobic interaction chromatography (HIC), the binding is independent of the surface hydrophobicity of the protein.
In hydrophobic interaction chromatography (HIC), the binding is independent of the surface hydrophobicity of the protein.
Proteins that do not bind a ligand in protein chromatography can be purified by altering the gene to express a fusion protein for affinity chromatography.
Proteins that do not bind a ligand in protein chromatography can be purified by altering the gene to express a fusion protein for affinity chromatography.
The elution order of peptides in size exclusion chromatography is primarily determined by their hydrophobicity.
The elution order of peptides in size exclusion chromatography is primarily determined by their hydrophobicity.
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Study Notes
- Chromatography is a technique used for analyzing or separating mixtures of substances, involving two distinct phases: stationary and mobile. (Oxford Dictionary of Science, 1999)
- Originating in the mid-1800s, chromatography was first documented for separating plant pigments. (Various sources)
- Chromatography has diverse applications, including forensics, food regulation, and athlete testing. (Various sources)
- Protein chromatography is a type of chromatography used for protein purification.
- Protein purification serves the purpose of preparing a pure protein sample to determine its properties or activity.
- Preliminary steps in protein purification include protein extraction and precipitation.
- Size-exclusion chromatography, ion-exchange chromatography, affinity chromatography, and hydrophobic interaction chromatography are methods used in protein purification. (Various sources)
- Chromatography techniques differ from HPLC/GC in terms of pressure, temperature, and flow rate.
- Protein extraction involves obtaining proteins from tissue or microbial cells, which necessitates breaking open the cells.
- Ion exchange chromatography separates proteins based on their charge by utilizing resins with oppositely charged groups.
- Size exclusion chromatography separates proteins based on their size.
- Affinity chromatography utilizes a specific ligand for binding the protein of interest.
- Chromatography techniques, such as size exclusion and affinity, require calibration for accurate results.
- Affinity chromatography utilizes a ligand covalently attached to the beads, and proteins bind to the beads based on their affinity for the ligand.
- Elution in affinity chromatography is achieved by using a high concentration of salt or a free ligand.
- Size exclusion and ion exchange chromatography are commonly used chromatographic methods.
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