What is antigen retrieval in immunofluorescence?
What is antigen retrieval in immunofluorescence?
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).
How does antigen retrieval work?
Antigen retrieval enables an antibody to access the target protein within the tissue. Masked epitopes can be recovered using either enzymatic/proteolytic antigen retrieval, or heat-induced antigen retrieval methods. In the enzymatic method, proteases such as proteinase K, trypsin, and pepsin are used.
What do you mean by antigen retrieval?
Antigen retrieval refers to any technique in which the masking of an epitope is reversed and epitope-antibody binding is restored.
Is antigen retrieval necessary in immunofluorescence?
Antigen retrieval is necessary to restore epitope-antibody reactivity. There are two main methods of antigen retrieval: Protease-Induced Epitope Retrieval (PIER) and Heat-Induced Epitope Retrieval (HIER). As with any technique, both methods must be optimized prior to any application.
How will you visualize an antigen in immunofluorescence assay?
1.1 Immunofluorescence Assay. IFA is a microscopic method that can detect and visualize the viral proteins expressed in cells via antigen—antibody reaction (Fig. 4.2). To visualize the antigen, the cells are then fixed by paraformaldehyde so that the proteins in the cells are poised to antibody binding.
What are the two types of immunofluorescence?
There are two classes of immunofluorescence techniques, primary (or direct) and secondary (or indirect).
Which type of immunofluorescent antibody test looks for antigen in the sample?
Indirect immunofluorescence detects the presence of antigen-specific antibodies in patient sera. The fluorescent antibody binds to the antigen-specific antibody rather than the antigen.
