Abstract
Heterotrimeric guanine nucleotide-binding proteins (G-proteins) play a pivotal role in a wide range of signal transduction pathways involved in multiple brain functions. Agonist-induced guanosine-5′-O-(3-[35S]thio)triphosphate ([35S]GTPγS) binding assay using filtration techniques has been widely utilized to investigate receptor-mediated activation of G-proteins. Since this method is applicable preferentially to Gi/o proteins, much effort has been made to capture receptor-mediated activation of other G-protein subclasses, such as Gs and Gq/11. Here, the method of [35S]GTPγS binding assay combined with immunoprecipitation using magnetic beads coated with anti-Gα antibody is described in detail. Some successful examples using this method in mammalian brain membranes are given, with several points to be noted when the reader employs this assay. The advantages and limitations of this method are also mentioned, and future direction in the application of this approach is indicated.
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Odagaki, Y. (2019). Guanosine-5′-O-(3-[35S]thio)triphosphate ([35S]GTPγS) Binding/Immunoprecipitation Assay Using Magnetic Beads Coated with Anti-Gα Antibody in Mammalian Brain Membranes. In: Odagaki, Y., Borroto-Escuela, D. (eds) Co-Immunoprecipitation Methods for Brain Tissue . Neuromethods, vol 144. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8985-0_8
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DOI: https://doi.org/10.1007/978-1-4939-8985-0_8
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