Abstract
High-quality antibodies must have high specificities and titers. The development and application of high-quality antibodies have greatly contributed to our current understanding of the life sciences, including neuroscience. However, it is often difficult to develop high-quality antibodies for neural proteins with complex and highly ordered structures and compositions, such as receptors and ion channels. As a researcher who has tackled the production of polyclonal antibodies, this chapter introduces the procedures for antigen peptide preparation, immunization, serum preparation, affinity purification, and specificity tests that I have adopted and modified over three decades. I believe that the two key points in high-quality antibody production are the whole-or-short strategy in which the full length or short peptides are preferable to the incomplete length as immunizing peptides and the gold-dust-fishing strategy by which useful antibodies are purified using affinity peptides designed to be shorter than, or displaced from, immunizing peptides.
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Acknowledgments
I appreciate all my colleagues who have used my antibodies in their splendid anatomical experiments.
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Watanabe, M. (2021). Production of High-Quality Antibodies for the Study of Receptors and Ion Channels. In: Lujan, R., Ciruela, F. (eds) Receptor and Ion Channel Detection in the Brain. Neuromethods, vol 169. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1522-5_1
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DOI: https://doi.org/10.1007/978-1-0716-1522-5_1
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