Abstract
Epitope tagging is a technique widely used in molecular and cellular biology. FLAG (DYKDDDDK), influenza virus hemagglutinin (YPYDVPDYA), and c-myc (EQKLISEEDL) tags are famous and are frequently used because high-affinity antibodies against these tags are commercially available. This technique is extremely useful for G protein-coupled receptor (GPCR) research because it is generally difficult to establish specific monoclonal antibodies (mAb) against GPCRs. We unexpectedly established a novel anti-FLAG mAb (2H8) during an attempt to generate an anti-mouse leukotriene B4 receptor 1 mAb. This mAb is a powerful tool to analyze various FLAG-fusion proteins, particularly GPCRs, both in vitro and in vivo. In this chapter, we describe experimental protocols to utilize the 2H8 mAb for flow cytometric, immunofluorescence staining, and immunoprecipitation analyses of various FLAG-tagged GPCRs.
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Sasaki, F., Yokomizo, T. (2015). A Novel Anti-FLAG Monoclonal Antibody Is Useful to Study GPCRs. In: Yokomizo, T., Murakami, M. (eds) Bioactive Lipid Mediators. Springer, Tokyo. https://doi.org/10.1007/978-4-431-55669-5_29
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DOI: https://doi.org/10.1007/978-4-431-55669-5_29
Publisher Name: Springer, Tokyo
Print ISBN: 978-4-431-55668-8
Online ISBN: 978-4-431-55669-5
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