Abstract
Immunohistochemistry is a powerful technique for determining both the presence of and the subcellular location of proteins within tissues. Zebrafish are particularly amenable to this technique and it is possible to localize proteins both in whole embryos and larvae, as well as sectioned material (Fig. 1). In this chapter, I will describe the basic technique developed for wholemount labeling of zebrafish embryos and variations that are required in specific situations. In addition, for each technique, pitfalls will be highlighted and ways to avoid these suggested.
Immunohistochemistry of wholemount and sectioned zebrafish tissue. (A) 28-h wholemount zebrafish embryo labeled with an antibody which recognizes the transcription factor, Pax6 (brown label) showing a lateral view with anterior to the left. (B) The 3-d larvae labeled with anti-acetylated tubulin antibody (brown), lateral view with anterior to the left. (C) Cross-section through a 3-d larvae eye labeled with anti-Pax6 antibody (brown) and a cone-specific antibody, fRet43 (black). (D) Oblique section through the surface of the optic nerve of an adult zebrafish labeled with anti- Pax2 antibody in yellow and anti-cytokeratin antibody in green and visualized using confocal microscopy. This preparation identified these Pax2 labelled cells as reticular astrocytes (1). (C is reproduced from ref. 2; D is reproduced from ref. 1.)
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© 1999 Humana Press Inc.
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Macdonald, R. (1999). Zebrafish Immunohistochemistry. In: Guille, M. (eds) Molecular Methods in Developmental Biology. Methods in Molecular Biology™, vol 127. Humana Press. https://doi.org/10.1385/1-59259-678-9:77
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DOI: https://doi.org/10.1385/1-59259-678-9:77
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