Separation techniques Flashcards
What is a centrifuge used for?
A centrifuge can be used to separate substances of differing density.
What happens in a centrifuge?
The more dense components settle in the form of a pellet, and the less dense components remain in the supernatant.
What are the different types of chromatography?
- paper chromatography
- thin layer chromatography
- affinity chromatography
What are paper and thin layer chromatography used for?
Paper and thin layer chromatography can be used for separating different substances such as amino acids and sugars.
What does the speed that each solute travels along the chromatography depend on?
The speed that each solute travels along the chromatography depends on the differing solubility in the solvent used.
What is affinity chromatography used for?
Affinity chromatography is used to separate proteins
What is the principle of affinity chromatography?
A solid matrix or gel column is created with specific molecules bound to the matrix or gel. Soluble target proteins in a mixture with a high affinity for these molecules become attached to them as the mixture passes down the column. Other non-target molecules with a weaker affinity are washed out
What is gel electrophoresis used for?
Gel electrophoresis is used to separate proteins and nuclei acids.
What is the principle of gel electrophoresis?
Charged macromolecules move through an electric field applied to a gel matrix
What are the two types of gel electrophoresis?
Native gels and SDS-PAGE
How do native gels separate proteins?
Native gels separate proteins by their shape, size and charge.
How does SDS-PAGE separate proteins?
SDS-PAGE separates proteins by size alone. This is because the SDS-PAGE gives all the molecules an equally negative charge and denatures them.
How else can proteins in a mixture be separated?
Proteins can be separated from a mixture using their isoelectric points (IEPs) If the solution is buffered to a specific pH, only the protein(s) that have an IEP of that pH will precipitate
How else can proteins be separated using their IEPs?
Proteins can also be separated using their IEPs in electrophoresis
