Phosphoprotein Detection by High-Throughput Flow Cytometry

Part of the Methods in Molecular Biology book series (MIMB, volume 1355)

Abstract

Phospho flow cytometry is a powerful technique for the detection of protein phosphorylation events that, like Western blotting, relies on phospho-epitope-specific antibodies. In contrast to the latter, however, multidimensional and directly quantifiable data is obtained at the single-cell level allowing separate analysis of small cell populations in complex cellular mixtures. Furthermore, up to 30 phospho-specific antibodies or antibodies identifying other posttranslational modifications in combination with cell surface markers can be analyzed in a single experiment. Utilizing a technique called fluorescent cell barcoding that enables combination of up to 64 samples into one tube for multiplex analysis and later data deconvolution, phospho flow cytometry is turned into a medium- to high-throughput technology.

Key words

Flow cytometry Phospho flow Phospho-specific antibody Fluorescent cell barcoding Signaling 

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Biotechnology CentreUniversity of OsloOsloNorway
  2. 2.K.G. Jebsen Centre for Cancer ImmunotherapyUniversity of OsloOsloNorway
  3. 3.Centre for Molecular Medicine Norway, Nordic EMBL PartnershipUniversity of OsloOsloNorway
  4. 4.K.G. Jebsen Inflammation Research CentreUniversity of OsloOsloNorway
  5. 5.Department of Infectious DiseasesUniversity of OsloOsloNorway

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