Protocols for Prostaglandin Receptor Localization in Eye Tissues Using Immunohistochemistry

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This chapter describes the detailed protocols for light and electron microscopic immunolocalization of prostaglandin (PG) receptors in ocular cells and tissues using immunofluorescence labeling of frozen sections, immunoperoxidase labeling of paraffin sections, and immunogold labeling of ultrathin sections. Unlike Western blotting analysis, immunohistochemistry allows for fine mapping of protein localization in intact tissues and cells, including assessment of relative abundance using image analysis software programs. Precise receptor subtype distribution is important for understanding mechanisms of action of therapeutic PG analogs, for identifying targets for novel treatments using subtype-specific high-affinity receptor agonists, and for improving safety profiles and minimizing potential side effects of anti-glaucoma pharmacotherapies.