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Questions and Answers
What is the main principle behind ultracentrifugation?
What is the main principle behind ultracentrifugation?
What term describes the liquid remaining above the precipitate after ultracentrifugation?
What term describes the liquid remaining above the precipitate after ultracentrifugation?
In electrophoresis, what factor primarily determines the movement of molecules through the solid support?
In electrophoresis, what factor primarily determines the movement of molecules through the solid support?
What is the purpose of staining the separated bands in plasma proteins electrophoresis?
What is the purpose of staining the separated bands in plasma proteins electrophoresis?
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Which method of protein separation involves applying an electric field across a gel?
Which method of protein separation involves applying an electric field across a gel?
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What happens to gammaglobulin levels during chronic infections?
What happens to gammaglobulin levels during chronic infections?
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What is the significance of the isoelectric point (pI) of a protein?
What is the significance of the isoelectric point (pI) of a protein?
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In chromatography, what determines the separation of a mixture's constituents?
In chromatography, what determines the separation of a mixture's constituents?
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What occurs during polyacrylamide gel electrophoresis (PAGE) with respect to protein size?
What occurs during polyacrylamide gel electrophoresis (PAGE) with respect to protein size?
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What impact does liver cirrhosis have on albumin levels?
What impact does liver cirrhosis have on albumin levels?
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Study Notes
Medical Biochemistry for Physiotherapy 2024-2025
- This course covers Medical Biochemistry for Physiotherapy students in 2024-2025.
Methods of Protein Separation
- Proteins are different physically and chemically.
- Methods for separating and studying proteins include Ultracentrifugation, Electrophoresis, and Chromatography.
Ultracentrifugation
- Definition: A mechanical process using centrifugal force to separate particles from a solution.
- Ultracentrifugation is high-speed centrifugation used for purifying macromolecules.
- Principle: Particles with different masses settle at different rates in a centrifuge.
- Heavier components settle quickly (precipitate), lighter components remain above (supernatant).
- Depends on molecular mass.
Electrophoresis
- Definition: A separation method of charged particles in an applied electric field.
- "Electro-" means electricity, "phoresis" means migration.
- Principle: Samples are placed on a solid support (e.g., gel, membrane) soaked in a conducting solution. An electric field is applied, causing charged molecules to migrate through the support.
- Depends on molecular mass and charge.
- Applications include Plasma Proteins Electrophoresis, Polyacrylamide Gel Electrophoresis, and Isoelectric Focusing.
Polyacrylamide Gel Electrophoresis (PAGE)
- Gel is cast vertically between glass plates.
- Samples are added, and an electric current is applied.
- Proteins migrate through the pores in the gel.
- Smaller proteins migrate faster.
- Larger proteins migrate slower.
Isoelectric Focusing (IEF)
- PAGE is set at different pH values to generate a pH gradient.
- Positively and negatively charged proteins move to the negative and positive electrodes, respectively, until they reach their isoelectric points (pI).
- Isoelectric point (pI): The pH at which a molecule carries no net electrical charge.
Chromatography
- Definition: A laboratory technique separating a sample dissolved in a mobile phase (gas or solvent) which moves through a stationary phase.
- Principle: Different mixture constituents have different affinities for the stationary phase, according to its nature.
- Depending on interactions, constituents travel at different velocities in the mobile phase, separating them.
- Depends on molecular mass and charge.
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Description
Explore the essential methods of protein separation in this Medical Biochemistry quiz designed for Physiotherapy students in the academic year 2024-2025. Learn about techniques such as Ultracentrifugation, Electrophoresis, and Chromatography. Test your knowledge on the principles and applications of these methods.
