Immunolocalization of Proteins in Situ by Light and Electron Microscopy
Part of the NATO ASI Series book series (NSSA, volume 135)
It is often desirable to localize proteins in situ in tissues and cells. Not only are we interested in the subcellular and tissue distribution of proteins, at times, the quantity in which a protein is present can be so low that its isolation and characterization is very difficult and tedious. For the histological identifaction of proteins, the most specific reagents are antibodies raised to it or to a closely related crossreacting immunogen — be they mono- or polyclonal. As a prerequiste, the specificity of the antibodies has to be ascertained by standard immunological methods, such as immunoblotting, enzyme immunoassay, or activity inhibition tests. In the following, basic technical recommendations for beginners in the field are given. They include:
Choice of Methods, Preparation of Specimen.
Antibodies and Antibody-markers.
Methods of staining.
KeywordsAnti Body Gold Chloride Unlabelled Antibody Drastic Method Antibody Penetration
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Major sources used for this overview
- 1).Bullock, G.R. & Petrusz, P. eds (1982–1985) Techniques in Immuno- cytochemistry Vol. 1–3, Academic Press.Google Scholar
- 2).Kuhlman, W.D. (1984) Immuno Enzyme Techniques in Cytochemistry, Verlag Chemie, Weinheim.Google Scholar
- 3).Marchalonis, J.J. & Warr, G.W. eds (1982) Antibody as a Tool. John Wiley & Sons Ltd., Chicester.Google Scholar
- 4).Sternberger, L. (1979) Immunocytochemistry, 2nd ed. John Wiley & Sons Ltd, New York.Google Scholar
© Plenum Press, New York 1987