1) c) separation techniques Flashcards
what does a centrifuge do?
Centrifuge separates substances of different densities.
More dense components settle in the pellet; less dense components remain in the supernatant.
what can paper and thin layer chromatography be used for?
Paper and thin layer chromatography can be used for separating different substances such as amino acids and sugars
The speed in which each solute travels along the chromatogram depends on its ________ __________ in the _______ _______
Differing solubility in the solvent used
The speed that each solute travels along the chromatogram depends on its differing solubility in the solvent used.
Describe the principles of affinity chromatography and how it separates proteins
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.
Principle of gel electrophoresis and its use in separating proteins and nucleic acids
Charged macromolecules move through an electric field applied to a gel matrix.
SEPERATES BASED OF CHARGE!
Describe the process of SDS Page
Separates proteins by size alone.
SDS–PAGE gives all the molecules an
equally negative charge and denatures them, separating proteins by size alone.
this means that the proteins can be separated based on their size as they migrate towards the positive electrode. Small proteins travel further through the gel than large proteins.
Proteins can be separated from a mixture using their _______________
Iso-electric Point
What is an Iso-electric point?
IEP is the pH at which a soluble protein has no net charge and will precipitate out of solution.
In relation to Iso-electric Point, If the pH has been buffered to a certain pH what proteins will precipitate?
If the solution is buffered to a specific pH, only the protein(s) that have an IEP of that pH will precipitate.
What is another way that proteins can be separated using their isoelectric points
Proteins can be separated using their isoelectric points in electrophoresis
How can proteins be separated using their isoelectric points in electrophoresis
Soluble proteins can be separated using an electric field and a pH gradient. A protein stops migrating through the gel at its IEP in the pH gradient because it has no net charge.
