1.1c - Separation Techniques Flashcards
What is a centrifuge used for?
to allow substances to be separated according to their density
More dense components do what?
settle in a pellet
Less dense components do what?
remain in the supernatant
Paper and thin layer chromatography can be used for…
separating substances such as amino acids and sugars
The speed that each solute travels along the chromatogram depends on…
its differing solubility in the solvent used
Affinity chromatography relies on…
the binding interaction between a protein and another substance (ligand)
A solid matrix or gel column is created with…
specific molecules bound to the matrix or gel
Soluble, target proteins in a mixture, with high affinity for their molecules become…
attached to them as the mixture passes down the column
Other non-target molecules with weaker affinity are…
washed out
Gel electrophoresis uses…
currents flowing through a buffer to separate proteins and nucleic acids
Charged macromolecules move through an…
electric field applied to a gel matrix
Native gels separate proteins by their… and why do they do this?
shape, size and charge because native gels do not denature the molecule
SDS-PAGE separates proteins by… and why?
size because it gives all the molecules an equally negative charge and denatures them
Proteins can be separated from a mixture using their what?
isoelectric points (IEP)
IEP is the pH…
at which a soluble protein has no net charge and will precipitate out of a solution
