Detecting recombinant proteins

Question 1

Describe electrophoresis

Answer 1

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

Question 2

Describe PAGE

Answer 2

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

Question 3

Describe SDS PAGE

Answer 3

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

Question 4

What are some ways to purify proteins?

Answer 4

Column chromatography
Size exclusion chromatography
Ion exchange chromatography
Affinity chromatography

Question 5

What is affinity chromatography based on?

Answer 5

Antibody:antigen
Enzyme:inhibitor