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MCAT Biochem > Protein Isolation > Flashcards

Flashcards in Protein Isolation Deck (15)
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1

Electrophoresis

uses a gel matrix to observe the migration of proteins in response to an electric field

2

Native PAGE

maintains the proteins shape but results are difficult to compare because the mass-to-charge ratio differs for each protein

3

SDS-PAGE

denatures the proteins and masks the native charge so that comparison of size is more accurate but the functional protein cannot be recaptured from the gel

4

Isoelectric Focusing

separates proteins by their isoelectric point (pI); the protein migrates toward na electrode until it reaches a region of the gel where pH = pI of the protein

5

Chromatography

separates protein mixtures on the basis of their affinity for a stationary phase or a mobile phase

6

Column chromatography

uses beads of a polar compound, like silica or alumina (stationary phase), with a non polar solvent (mobile phase)

7

Ion-exchange chromatography

uses a charged column and a variably saline eluent

8

Size exclusion chromatography

relies on porous beads

Larger molecules elute first because that are not trapped in the small pores

9

Affinity chromatography

uses a bound receptor or ligand and an fluent with free ligand or a receptor for the protein of interest

10

Homogenization

crushing, grinding, or blending the tissue of interest into an evenly mixed solution

11

Centrifugation

isolates the proteins from much smaller molecules before other isolation techniques must be employed

12

SDS detergent disrupts

all noncovalent interactions

13

retention time

the amount of time a solute remains in the stationary phase

14

what can be triggered to help elute the protein of interest in a column chromatography?

solvent polarity
pH
salinity

15

2 drawbacks of using the affinity chromatography

1. the protein of interest may not elute from the column because its affinity is too high
2. it may be permanently bound to the free receptor in the eluent