[20] Anti-fusion protein antibodies specific for receptor subtypes

Publisher Summary

This chapter discusses anti-fusion protein antibodies specific for receptor subtypes. Immunocytochemistry of specific neurochemical markers has been very useful for the study of functional neuroanatomy and has been employed for the localization of presynaptic markers (such as neuropeptides, neurotransmitters, and their synthetic enzymes) of chemically defined neurons. Several additional aspects of immunochemical detection provide distinct advantages over radioligand-binding autoradiography for the localization of neurochemical markers. First, whereas binding assays depend on a high-affinity radioligands, antibodies can be generated against many different types of molecules (e.g., receptors, transporters, synthetic enzymes, neurotransmitters, or neuropeptides), regardless of their ligand-binding properties. Second, spatial resolution at the light and electron microscopic levels is superior with immunocytochemical techniques, permitting more precise neuroanatomical localization and allowing more specific questions to be addressed. Third, immunocytochemical methods are readily adapted to multiple-label experiments, using immunofluorescent, silver-intensified immunogold, or immunoperoxidase methods. Molecular biological techniques can be used to raise antibodies against select regions of a receptor subtype. This process avoids several problems with antigen purification, as used in conventional immunological schemes, and produces greater antibody specificity.