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CD95 pp 173-188 | Cite as

Site-Specific Detection of Tyrosine Phosphorylated CD95 Following Protein Separation by Conventional and Phospho-Protein Affinity SDS-PAGE

  • Krittalak Chakrabandhu
  • Sébastien Huault
  • Anne-Odile HueberEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1557)

Abstract

Phosphorylation of two tyrosines in the death domain of CD95 is a critical mechanism in determining the receptor’s choices between cell death and survival signals. Recently, site-specific monoclonal antibodies against phosphorylated tyrosines of CD95 have been generated and used to successfully detect each phosphorylated death domain tyrosine of CD95 directly and separately by immunoblotting. Here we provide detailed protocols and useful tips for a successful site-specific detection of phosphorylated death domain tyrosine of CD95 following a protein separation by sizes (conventional SDS-PAGE) and by degrees of phosphorylation (phospho-protein affinity, mobility shift SDS-PAGE).

Key words

SDS-PAGE Immunoblot Phos-tag™ Phosphorylation CD95 Mobility shift 

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Krittalak Chakrabandhu
    • 1
  • Sébastien Huault
    • 1
  • Anne-Odile Hueber
    • 2
    Email author
  1. 1.Université Côte d’Azur, Institut de Biologie Valrose, CNRS UMR 7277, Inserm UMR 1091Parc Valrose, Bâtiment des Sciences NaturellesNiceFrance
  2. 2.Institut de Biologie Valrose, CNRS UMR 7277, INSERM UMR,Université de NiceNiceFrance

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