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A Short Introduction to Immunocytochemistry and a Protocol for Immunovisualization of Proteins with Alkaline Phosphatase

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Methods in Plant Electron Microscopy and Cytochemistry

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Abstract

Immunocytochemical methods have been widely applied to visualize proteins, carbohydrates, or lipids in sectioned material. The advantage of using immunocytochemistry is to be able to localize the molecules of interest within the tissue. Several procedures have been described. Basically, these procedures can be split into four main steps that are described in subheadings: (1) tissue preparation, (2) the primary antibodies, (3) the visualization of the target, and (4) enhancement of signals with antibody complexes. In addition, a protocol for alkaline phosphatase will be presented in detail in Subheading 5. The terms “primary” and “secondary” antibodies refer to the order in which they are applied to the target. The immunocytochemical procedures are not limited to sectionedmaterial. The immunocytochemical procedures can also be used for fixed and enzymatically permeabilized cells of Saccharomyces cerevisiae (1,2), Schizosaccharomyces pombe (3), and Chara (4) protoplasts and organelles (5). The immunocytochemical techniques are used in combination with other techniques: flow cytometry (6), tissue prints (7), in situ hybridization (8–10), and in situ PCR (polymerase chain reaction) (11–13).

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Authors
  1. John E. Nielsen
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Editors and Affiliations

  1. Mary Baldwin College, Richmond, VA, USA

    William V. Dashek

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© 2000 Springer Science+Business Media New York

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Nielsen, J.E. (2000). A Short Introduction to Immunocytochemistry and a Protocol for Immunovisualization of Proteins with Alkaline Phosphatase. In: Dashek, W.V. (eds) Methods in Plant Electron Microscopy and Cytochemistry. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-59259-232-6_6

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  • DOI: https://doi.org/10.1007/978-1-59259-232-6_6

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61737-199-8

  • Online ISBN: 978-1-59259-232-6

  • eBook Packages: Springer Book Archive

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