Methods of Antigen Retrieval

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An antigen is protein that is used to start immune response in an animal. The animal reacts by producing antibodies to an individual epitopes or antigenic determinant region located on the antigen. Antigen has multiple epitopes which can generate antibodies (Dalcik et al, 2012). The antigen can be improved by pre-treatment with antigen retrieval reagent which break the protein cross links formed by formalin fixation thus expose unseen antigenic site. There are different kind of antigens like CD20, AE1, epithelial membrane antigen, MIB1, ER, CD43 and HMB45.

The immunohistochemistry takes advantage of reactive specificity which exists between an antibody and the corresponding antigen within tissue or cells. The antigen- antibody visualization complexes can be achieved through a number of colorimeteric reaction, typically using a fluorescently labelled antibody or immunoperoxidase reporting system. When tissue is formalin fixed and paraffin embedded this process sometimes results in the loss of antigenicity in the tissue. to overcome this problem many different methods have been developed to reconstitute antigenicity by gently heating the tissue sections once rehydrated in varius mild salt solution (D’ Amico et al. 2009; Leong et al. 2010; Shi et al.2011). The antigen retrieval methods cause the protein in the tissue to unfold and then refold with the possibility of having the unavailable epitope to now be exposed and available for antibody binding. (Onul et al. 2012)

There are many different methods can be used for antigen retrieval such in Room temperature epitope retrieval method like Hydrochloric acid methods and formic acid method is used. In heat induced epitope retrieval, methods such as citrate buffer (pH6), citrate-EDTA bu...

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...uclear immunostaining for Antigen retrieval. The study by Shi SR show the non-specific background was stronger and the reactivity of positive nuclear was weak compare to those obtained by antigen retrieval techniques. The study also shows the difference in solution used for antigen retrieval, the immunostaining intensity was satisfactory, especially in 5% urea solution and citric acid buffer, and in distilled water less intensity was observed.
The antigen retrieval technique is changing; there may be different advance or approach to improve immunostaining. The presence knowledge in specific approach differ for different antigen and different antibody for varies duration of fixation and also perhaps for different fixatives. The original antgen retrieval techniques is now widespread use and is important in organic chemical research into formalin fixed tissue section.

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