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Immunofluorescent Labelling of Enzymes

  • G. Schmid
  • H. Grisebach
Part of the Modern Methods of Plant Analysis book series (MOLMETHPLANT, volume 4)

Abstract

Besides knowledge of kinetic and regulatory behaviour of enzymes, the determination of their intra- and intercellular localization is an important prerequisite for understanding cellular metabolism. One of the most sensitive and specific techniques for reaching this goal is the in situ localization of enzymes by immunological methods. In principle the specificity of the antigen(enzyme)-antibody reaction is combined with a labelling technique which allows detection of the enzyme-antibody complex in a tissue section either by light or electron microscopy. For immunoelectron microscopy (Hoyer and Bucana 1982) ferritin, peroxidase, and gold are the most widely used markers for direct and indirect labelling. For example, nitrate reductase was localized in the cell wall-plasmalemma region and in the tonoplast membranes of Neurospora crassa cells by incubation with antiserum against nitrate reductase followed by an incubation with ferritin-labelled goat antirabbit IgG (indirect method) (Roldàn et al. 1982). Gold-labelled protein A (a cell wall protein from Staphylococcus aureus with a high affinity for the Fc fragment of IgG) was used to localize an extracellular protease of Nectria galligena in infected apple tissue (Rey and Noble 1984).

Keywords

Neurospora Crassa Immunofluorescent Labelling Cicer Arietinum Coniferyl Alcohol Filter Combination 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Springer-Verlag Berlin Heidelberg 1986

Authors and Affiliations

  • G. Schmid
  • H. Grisebach

There are no affiliations available

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