Immunocytochemical Localization of Receptors Using Light and Confocal Microscopy With Application to the Phenotypic Characterization of Knock-Out Mice
- 619 Downloads
This chapter describes the immunocytochemical (ICC) localization of receptors in tissue sections and cells growing in culture. The protocol is illustrated using examples of how site-directed polyclonal antisera can be used to distinguish between subtypes of G protein-coupled receptors (GPCRs; endothelin ETA and ETB), (1) in sections of human tissue or whole-body sections of mice, as well as mapping previously designated orphan receptors (oGPCR) recently paired with their cognate ligand illustrated by visualizing receptors for the peptide ghrelin (2). The methods are applicable to other receptor families.
KeywordsAlexa Fluor Primary Antiserum Secondary Antiserum Swine Serum Nonspecific Background Staining
- 13.Kurihara, H., Kurihara, J., and Yazaki, Y. (2001) Lessons from gene deletion of endothelin systems. Hdbk. Exp. Pharmacol. 152, 141–154.Google Scholar
- 16.Kuc, R. E. Immunocytochemical localization of endothelin peptides, precursors and endothelin-converting enzymes. Methods Mol. Biol. 206, 3–9.Google Scholar