How does heat induced antigen retrieval work?
Heat induced epitope retrieval (HIER) may be defined in the simplest terms as the use of heat coupled with specific buffered solutions to recover antigen reactivity in formalin fixed paraffin embedded tissue. The HIER procedure has had a huge impact on the way IHC is utilized in the diagnostic process.
What is antigen retrieval process?
Antigen retrieval is an effective method of unmasking antigenic epitopes on the surface of formalin-fixed paraffin-embedded (FFPE) tissue sections. The antigen retrieval technique breaks the methylene bridges between epitopes and unrelated proteins to expose antigenic sites for antibody binding (1) (Figure 1).
What is heat mediated antigen retrieval?
Heat-induced epitope retrieval is most often performed using a pressure cooker, a microwave, or a vegetable steamer. Some labs use a water bath set to 60°C and incubate the slides in retrieval solution overnight. This applies also to the choice of buffer used for heat-mediated retrieval.
How do you make an antigen retrieval solution?
Sodium citrate buffer (10 mM Sodium citrate, 0.05% Tween 20, pH 6.0)
- Tri-sodium citrate (dihydrate) 2.94 g.
- Distilled water 1 L.
- Mix to dissolve. Adjust pH to 6.0 with 1N HCl.
- Add 0.5 mL Tween 20 and mix well. Store at room temperature for 3 months or at.
- 4°C for longer storage.
What are the methods of antigen retrieval technique?
The two methods for antigen retrieval are heat induced epitope retrieval (HIER) and enzymatic retrieval.
What are the different methods of retrieval of masked antigens?
When discussing antigen retrieval methods, techniques generally fall into two main categories, protease-induced epitope retrieval (PIER) and heat-induced epitope retrieval (HIER).
How do you Deparaffinize sections?
a) Heat treatment (recommended method): Place slides in a container and cover with 10 mM sodium citrate buffer, pH 6.0; or with 50 mM glycine-HCl buffer (glycine: sc-29096), pH 3.5, with 0.01% (w/v) EDTA (EDTA: sc-29092). Heat at 95º C for 5 minutes.
Why is xylene used for Deparaffinization?
Histologists are usually exposed to xylene during histoprocessing, deparaffinization, and clearing prior to coverslipping. [10] Worldwide, xylene is considered to be the most commonly used dewaxing and clearing agent as it renders tissue transparent and removes alcohols from tissues rapidly.