Detecting recombinant proteins Flashcards Preview

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Flashcards in Detecting recombinant proteins Deck (5)
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1

Describe electrophoresis

Separation based on charge
8.3-8.6 pH
DNA moves towards the anode
Needs support medium ie starch/agarose

2

Describe PAGE

Uses polyacrylamide gel
e- flow is even
Mobility based on charge and gel pore size

3

Describe SDS PAGE

Sodium dodecyl sulfate unfolds the protein and coats it with a negative charge
Charge:mass the same for all proteins
Mobility based on mass
Sieving so small proteins are at the front

4

What are some ways to purify proteins?

Column chromatography
Size exclusion chromatography
Ion exchange chromatography
Affinity chromatography

5

What is affinity chromatography based on?

Antibody:antigen
Enzyme:inhibitor