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Multiparameter Flow Cytometry of Fluorescent Protein Reporters

  • Protocol
Flow Cytometry Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 263))

Abstract

Reporters based on the green fluorescent protein (GFP) from the jellyfish Aequorea victoria and GFP-like proteins from other marine organisms provide valuable tools to monitor gene transfer and expression noninvasively in living cells. Stable cell lines were generated from the Sp2/0-Ag14 hybridoma that express up to three spectral enhanced versions of GFP, the enhanced cyan fluorescent protein (ECFP), the enhanced green fluorescent protein (EGFP), and the enhanced yellow fluorescent protein (EYFP), and/or a variant of the Discosoma coral red fluorescent protein (DsRed). The panel of lines was used to demonstrate a flow cytometric procedure for simultaneous analysis of all four fluorescent proteins that utilizes dual-laser excitation at 488 nm and 407 nm. Additional schemes for simultaneous detection of two, three or four of these fluorescent proteins are also presented.

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Author information

Authors and Affiliations

  1. Flow Cytometry Facility and Hematopoiesis Department, Holland Laboratory, American Red Cross, Rockville, MD

    Teresa S. Hawley

  2. Verity Software House Inc., Topsham, ME

    Donald J. Herbert

  3. Healthspex, Inc., Knoxville, TN

    Shannon S. Eaker

  4. Hematopoiesis Department and Blood and Cell Therapy Development, Holland Laboratory, American Red Cross, Rockville, MD

    Robert G. Hawley

  5. Department of Anatomy and Cell Biology, The George Washington University, Washington, DC

    Robert G. Hawley

Authors
  1. Teresa S. Hawley
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  2. Donald J. Herbert
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  3. Shannon S. Eaker
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  4. Robert G. Hawley
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Editor information

Editors and Affiliations

  1. American Red Cross, Rockville, MD

    Teresa S. Hawley  & Robert G. Hawley  & 

  2. The George Washington University, Washington, DC

    Robert G. Hawley

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© 2004 Humana Press Inc.

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Hawley, T.S., Herbert, D.J., Eaker, S.S., Hawley, R.G. (2004). Multiparameter Flow Cytometry of Fluorescent Protein Reporters. In: Hawley, T.S., Hawley, R.G. (eds) Flow Cytometry Protocols. Methods in Molecular Biology™, vol 263. Humana Press. https://doi.org/10.1385/1-59259-773-4:219

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  • DOI: https://doi.org/10.1385/1-59259-773-4:219

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-234-6

  • Online ISBN: 978-1-59259-773-4

  • eBook Packages: Springer Protocols

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