La 8 & 9 - Affinity Chromatography of Concanavalin A (Con A)

Question 1

What is affinity chromatography?

Answer 1

A method used to separate mixtures by manipulating specific interactions between antigen and antibody, receptor and enzyme, or enzyme and substrate

Question 2

What is the purpose of affinity chromatography?

Answer 2

Purify and concentrate a substance in a mixtures

Question 3

What is concanavalin A (con A)?

Answer 3

Carbohydrate binding protein = Lectin

Question 4

How do you purify Con A?

Answer 4

Dextrose

Question 5

Where did the crude extract come from?

Answer 5

Jack bean meal extract

Question 6

Where did the effluent come from?

Answer 6

The wash with 1 M NaCl

Question 7

Where did the eluate come from?

Answer 7

Samples collected after 5 mL of 1 M NaCl/1M Dextrose was added

Question 8

How did we elute ConA from the column?

Answer 8

Dextrose

Question 9

How did we detect ConA on the membrane?

Answer 9

It binds to HRP to make purple color

Question 10

Results

Answer 10

Last Eluate > 2nd Eluate > ConA control> Jack bean > First Eluate > Effluent