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What is the main principle behind electrophoresis?
What is the main principle behind electrophoresis?
What is the purpose of assays to determine protein concentration?
What is the purpose of assays to determine protein concentration?
Which method relies on changes in the color of a compound upon binding to proteins to determine protein concentration?
Which method relies on changes in the color of a compound upon binding to proteins to determine protein concentration?
What is the significance of a darker purple color in the BCA assay reaction?
What is the significance of a darker purple color in the BCA assay reaction?
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How are macromolecules in a mixture separated during electrophoresis?
How are macromolecules in a mixture separated during electrophoresis?
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What is the purpose of comparing color changes in a spectrophotometric plate reader during a total protein assay?
What is the purpose of comparing color changes in a spectrophotometric plate reader during a total protein assay?
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What is the main purpose of using Fluorescence Resonance Energy Transfer (FRET) in protein studies?
What is the main purpose of using Fluorescence Resonance Energy Transfer (FRET) in protein studies?
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What happens if two fluorophores are distanced efficiently in FRET?
What happens if two fluorophores are distanced efficiently in FRET?
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Why is labeling two proteins that are already expected to interact not a good tool to discover new interactions using FRET?
Why is labeling two proteins that are already expected to interact not a good tool to discover new interactions using FRET?
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In the context of FRET, what does the emission of fluorescent light by one fluorophore do to the other fluorophore?
In the context of FRET, what does the emission of fluorescent light by one fluorophore do to the other fluorophore?
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What is the role of Fluorescence Resonance Energy Transfer (FRET) in detecting change in protein conformation?
What is the role of Fluorescence Resonance Energy Transfer (FRET) in detecting change in protein conformation?
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What can FRET be used to detect in live cells over time?
What can FRET be used to detect in live cells over time?
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What is the main purpose of two-dimensional gel electrophoresis (2D-PAGE)?
What is the main purpose of two-dimensional gel electrophoresis (2D-PAGE)?
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What can cause molecular supercomplexes to dissociate, leading to less effective respiratory chain and ATP depletion syndromes?
What can cause molecular supercomplexes to dissociate, leading to less effective respiratory chain and ATP depletion syndromes?
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Which technique is used to analyze protein modifications that cannot be detected by SDS-PAGE?
Which technique is used to analyze protein modifications that cannot be detected by SDS-PAGE?
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What is the main limitation of the Sanger method for protein sequencing?
What is the main limitation of the Sanger method for protein sequencing?
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How are proteins identified in cellular homogenate mixtures before extraction and digestion with trypsin?
How are proteins identified in cellular homogenate mixtures before extraction and digestion with trypsin?
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Which technique involves cleaving proteins between Lysine and Arginine residues to generate peptides for mass spectrometry analysis?
Which technique involves cleaving proteins between Lysine and Arginine residues to generate peptides for mass spectrometry analysis?
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Which method can be used to study protein-protein interactions by measuring the distance-dependent transfer of energy from a donor molecule to an acceptor molecule?
Which method can be used to study protein-protein interactions by measuring the distance-dependent transfer of energy from a donor molecule to an acceptor molecule?
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What is the traditional approach for solving the tertiary structure of proteins?
What is the traditional approach for solving the tertiary structure of proteins?
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Which method involves biotinylating proteins that come close to the protein of interest to track interaction partners of a protein?
Which method involves biotinylating proteins that come close to the protein of interest to track interaction partners of a protein?
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Which method is used to sequence peptide fragments and compare peptide masses with a protein database for protein identification?
Which method is used to sequence peptide fragments and compare peptide masses with a protein database for protein identification?
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What is the artifact that can occur in Co-immunoprecipitation involving non-specific binding and unspecific proteins stuck to beads, requiring controls to distinguish genuine interactions?
What is the artifact that can occur in Co-immunoprecipitation involving non-specific binding and unspecific proteins stuck to beads, requiring controls to distinguish genuine interactions?
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Which method examines the relative abundance of peptide masses to identify proteins, requiring nano-pico mole amounts of the protein?
Which method examines the relative abundance of peptide masses to identify proteins, requiring nano-pico mole amounts of the protein?
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What initiates the chemical crosslinking of Acrylamide and Bisacrylamide in PAGE gels?
What initiates the chemical crosslinking of Acrylamide and Bisacrylamide in PAGE gels?
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Which variant of PAGE linearizes proteins and destroys all chemical/covalent links, allowing proteins to become negatively charged and migrate based on size?
Which variant of PAGE linearizes proteins and destroys all chemical/covalent links, allowing proteins to become negatively charged and migrate based on size?
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Which type of PAGE preserves protein-protein interactions that exist in vivo and allows recovery of proteins in their functional form, with retained enzyme activity?
Which type of PAGE preserves protein-protein interactions that exist in vivo and allows recovery of proteins in their functional form, with retained enzyme activity?
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What does 2-Dimensional PAGE involve?
What does 2-Dimensional PAGE involve?
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What can be added to the gel in PAGE to detect the presence of a specific protein in a sample?
What can be added to the gel in PAGE to detect the presence of a specific protein in a sample?
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What is used in PAGE to compare migration distances of proteins of interest to molecular weight ladders and estimate their apparent molecular weight based on the migration distance?
What is used in PAGE to compare migration distances of proteins of interest to molecular weight ladders and estimate their apparent molecular weight based on the migration distance?
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Study Notes
Polyacrylamide Gel Electrophoresis (PAGE) in Biomedicine
- Polyacrylamide Gel Electrophoresis (PAGE) is a significant advancement in protein resolution and characterization.
- PAGE can be performed using various matrices such as paper and different gels for protein separation.
- PAGE gels are formed using chemical crosslinking of Acrylamide and Bisacrylamide, initiated by ammonium persulphate and the base TEMED, solidifying in 10-15 minutes.
- The percentage of acrylamide in the gels varies based on the nature and size of the macromolecule being separated, with lower percentages used for nucleic acids and higher percentages for proteins.
- SDS-PAGE (Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis) is the most common variant, which linearizes proteins and destroys all chemical/covalent links, allowing proteins to become negatively charged and migrate based on size.
- SDS-PAGE can be performed in reducing or non-reducing conditions to examine associations between proteins, with reducing conditions using agents that reduce disulfide bonds.
- Native PAGE preserves protein-protein interactions that exist in vivo and allows recovery of proteins in their functional form, with retained enzyme activity.
- 2-Dimensional PAGE involves two successive rounds of electrophoresis and dramatically improves separations and resolution of complex mixtures of proteins.
- Gradients can be used in PAGE to improve separation, with injection wells and changing pore sizes in the gel.
- PAGE allows the resolution of proteins in complex mixtures, estimation of their size, and determination of some of their properties.
- Specific antibodies can be added to the gel to detect the presence of a specific protein in a sample, allowing for precise measurements.
- PAGE gels can be used to compare migration distances of proteins of interest to molecular weight ladders and estimate their apparent molecular weight based on the migration distance.
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Test your knowledge of Polyacrylamide Gel Electrophoresis (PAGE) in biomedicine with this quiz. Explore the different types of PAGE, its applications, and the techniques used for protein separation and characterization.
