Multiple antigen immunostaining

Key words Multiple antigen immunostaining, Immunoenzymatic methods, Immunofluorescence,... 

The most common fluorochromes used for multiple antigen immunostaining... 

Krenacs, X. Krenacs, L. Raffeld, M. Multiple antigen immunostaining procedures. Methods Mol. Biol. 2010,588, 281-300. 

Labeling with fluorescent phalloidins may be combined with immunostaining. In this case, the phalloidin-staining solution can be applied in a mixture with fluores-cently labeled secondary antibodies. Combination of immunostaining with fluorescent phalloidins and fluorescently counterstained nuclei are extremely useful in multiple labeling strategies to locate antigens of interest with specific components of the cell. 

Another method takes advantage of significant differences in detection sensitivities of two immunostaining procedures. For example, the streptavidin-biotin method with biotinylated-tyr-amide amplification can detect antigens at such a low concentration of the primary antibody, that they can t be detected with the traditional indirect methods used in the subsequent immunoreac-tions (38, 39). The principles of double/multiple immunostaining techniques are summarized in Table 3. 

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