Antigen retrieval

Question 1

Purpose of antigen retrieval

Answer 1

To increase the accessibility of antibody to antigen

Question 2

General methods of antigen retrieval: (2)

Answer 2

1. Proteolytic Enzyme Retrieval/ Protease Induced Epitope Retrieval (PIER)
2. Heat Induced Epitope Retrieval (HIER)

Question 3

Most common enzymes used in PIER: (3)

Answer 3

1. Trypsin
2. Protease

Question 4

Why are trypsin or protease the most common used enzyme in PIER?

Answer 4

They can break/ remove formalin crosslinks

Question 5

Techniques in HIER: (7)

Answer 5

1. Microwave antigen retrieval (20 min)
2. Pressure cooker antigen retrieval (less
   time consuming)
3. Autoclave heating
4. Water bath heating
5. Steamer heating
6. Decloaker heating
7. Combination of microwave & enzyme
   digestion