Unit 1 - Topic 1 - Laboratory Techniques for Biologists - Section C - Separation techniques Flashcards

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1
Q

What is a centrifuge used for?

A

Centrifuge are used to separate substances of differing density

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2
Q

What are the names for the separated components?

A

More dense components settle in the pellet; less dense components remain in the supernatant.

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3
Q

What can be used for separating different substances such as amino acids and sugars?

A

Paper and thin layer chromatography

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4
Q

What does solubility in the solvent effect?

A

The speed that each solute travels along the chromatogram

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5
Q

Explain affinity chromatography and its use in separating proteins

A
  • 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
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6
Q

What does gel electrophoresis separate?

A

Proteins and nucleic acids

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7
Q

How does gel electrophoresis work

A

Charged macromolecules move though an electric field applied to a gel matrix

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8
Q

What do native gels separate proteins by

A

Shape, size and charge

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9
Q

What do native gels separate proteins by and how do they do this

A

Shape, size and charge. They do not denature the molecules

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10
Q

How does SDS-PAGE separate proteins

A

SDS–PAGE gives all the molecules an equally negative charge and denatures them, separating proteins by size alone.

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11
Q

What is an IEP and what does it separate

A

The isoelectric point separates proteins. An IEP is the pH at which a soluble protein has no net charge and will precipitate out of solution.

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12
Q

How does a buffer effect IEP

A

If the solution is buffered to a specific pH, only the protein(s) that have an IEP of that pH will precipitate

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13
Q

Proteins can also be separated using their IEPs in electrophoresis how is this done

A

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.

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