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Questions and Answers
What is the primary structure of proteins?
What is the primary structure of proteins?
What is the isoelectric point (pI) of an amino acid?
What is the isoelectric point (pI) of an amino acid?
What are peptides?
What are peptides?
How many common amino acids are there?
How many common amino acids are there?
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What are proteins' various biological functions?
What are proteins' various biological functions?
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What is the zwitterion form of an amino acid?
What is the zwitterion form of an amino acid?
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What did Dutch chemist GJ Mulder observe while investigating substances in milk and eggs?
What did Dutch chemist GJ Mulder observe while investigating substances in milk and eggs?
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Who suggested that substances in milk and eggs should be called proteins?
Who suggested that substances in milk and eggs should be called proteins?
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What is the topic of Week 9 in the course schedule?
What is the topic of Week 9 in the course schedule?
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Who are the authors of the book 'Bioanalytical Chemistry' mentioned in the text?
Who are the authors of the book 'Bioanalytical Chemistry' mentioned in the text?
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What is the main focus of Week 11 in the course schedule?
What is the main focus of Week 11 in the course schedule?
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What did Swedish chemist JJ Berzelius suggest about the substances in milk and eggs?
What did Swedish chemist JJ Berzelius suggest about the substances in milk and eggs?
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What type of bonds stabilize α-helix structure in peptide chains?
What type of bonds stabilize α-helix structure in peptide chains?
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What is the primary stabilizing force in β-pleated sheet structure?
What is the primary stabilizing force in β-pleated sheet structure?
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In which protein structure is the polypeptide chain reversed in direction?
In which protein structure is the polypeptide chain reversed in direction?
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What type of bonds are involved in forming tertiary protein structure?
What type of bonds are involved in forming tertiary protein structure?
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What technique is used to study different levels of protein structure by measuring the absorption of circularly polarized light?
What technique is used to study different levels of protein structure by measuring the absorption of circularly polarized light?
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What is the primary aim of protein purification?
What is the primary aim of protein purification?
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What is the wavelength at which proteins are detected in the Lowry method?
What is the wavelength at which proteins are detected in the Lowry method?
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Which protein quantification method is known for being 100 times more sensitive than the Biuret method?
Which protein quantification method is known for being 100 times more sensitive than the Biuret method?
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What type of electrophoresis separates proteins based on their net charge and size?
What type of electrophoresis separates proteins based on their net charge and size?
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Who pioneered protein sequencing and determined the sequence of bovine insulin?
Who pioneered protein sequencing and determined the sequence of bovine insulin?
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What gel staining method incorporates SDS to fully denature proteins and dissociate them?
What gel staining method incorporates SDS to fully denature proteins and dissociate them?
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What is the purpose of dye binding methods like the Bradford protein assay?
What is the purpose of dye binding methods like the Bradford protein assay?
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Which method of protein extraction is specifically mentioned as being hand-operated or motor-driven?
Which method of protein extraction is specifically mentioned as being hand-operated or motor-driven?
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What is the purpose of using detergents in protein extraction?
What is the purpose of using detergents in protein extraction?
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Which substance is recommended to protect thiol groups from oxidation during protein purification?
Which substance is recommended to protect thiol groups from oxidation during protein purification?
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What is the purpose of using chelators in protein purification?
What is the purpose of using chelators in protein purification?
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What is the primary factor that influences protein precipitation using organic solvents?
What is the primary factor that influences protein precipitation using organic solvents?
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Which chromatographic step involves considerations for sample volume, protein concentration, and efficiency of separation methods?
Which chromatographic step involves considerations for sample volume, protein concentration, and efficiency of separation methods?
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What is the purpose of using detergents in protein extraction?
What is the purpose of using detergents in protein extraction?
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What is the function of reducing agents like DTE, DTT, and mercaptoethanol during protein purification?
What is the function of reducing agents like DTE, DTT, and mercaptoethanol during protein purification?
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How does ammonium sulphate salting out depend on protein properties?
How does ammonium sulphate salting out depend on protein properties?
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What is the purpose of using chelators in protein purification?
What is the purpose of using chelators in protein purification?
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What is the primary function of protein separation by gel filtration chromatography?
What is the primary function of protein separation by gel filtration chromatography?
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How do organic solvents induce protein precipitation?
How do organic solvents induce protein precipitation?
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Study Notes
Protein Purification Techniques and Considerations
- Protein extraction methods include hand-operated or motor-driven blenders, ultrasound, vibrating bead mill, and Manton-Gaulin homogenizer
- Factors to consider in extraction medium include temperature (2-8°C) and time, with mild, moderate, and vigorous techniques available
- Extraction medium considerations include buffer salts, pH, detergents, reducing agents, chelators, proteolytic inhibitors, and bacteriostatics
- Detergents are used to break hydrophobic interactions, with concentrations below critical micelle concentration recommended
- Reducing agents like DTE, DTT, and mercaptoethanol protect thiol groups from oxidation during purification
- Chelators and metal ions may stabilize proteins but heavy metal ions can enhance oxidation; EDTA can be used to remove heavy metals
- Proteolytic inhibitors and bacteriostatics are used to prevent protein degradation and bacterial growth, respectively
- Protein precipitation methods include salting out with ammonium sulphate, organic solvents, and dialysis through semi-permeable membranes
- Ammonium sulphate salting out depends on protein hydrophobicity and competition for water molecules
- Organic solvents induce protein precipitation by reducing water activity and breaking protein intramolecular interactions
- Dialysis is used to separate proteins from small molecules and remove salts and ammonium sulphate
- Protein separation by chromatographic steps includes gel filtration, ion exchange, and affinity chromatography, with considerations for sample volume, protein concentration, and efficiency of separation methods
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Description
Test your knowledge of protein quantification, analysis, and purification techniques with this quiz. Explore methods such as UV absorption, electrophoresis, protein sequencing, and extraction techniques including protein precipitation and chromatography. Gain insights into factors to consider in extraction medium, detergents, reducing agents, and chromatographic separation methods.
