Abstract
Publisher Summary This chapter describes the methods for direct and indirect immunoperoxidase staining of S-100 proteins and calmodulin in tissue sections and cultured cells. Preparation of peroxidase-labeled antibodies is now widely used for light and electron microscopic immunocytochemistry, and is also applicable to stain a specific antigen on nitrocellulose sheets transferred from polyacrylamide gels. The immunoperoxidase method exhibits the advantage of the stability of the enzyme reaction product, the easiness in the counter staining by the conventional staining methods, and usefulness in the immunoelectron microscopic study. The immunofluorescent method often provides a higher resolution at the light microscopic level. For visualization of antibody molecules bound to antigen by the immunoperoxidase method, direct staining (using peroxidase directly conjugated to specific antibody) or indirect staining (in which tissue or cells are reacted with the specific antibody followed by reaction with peroxidaselabeled second 3,-globulin) is employed. There are some modifications of the indirect staining method, such as the peroxidase-anti-peroxidase (PAP) method, in which PAP complex (rabbit anti-peroxidase and free peroxidase) is linked with the primary rabbit antigen-antibody complex by the addition of goat-anti-rabbit IgG antibody.
Published Version
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