Determining the Phosphorylation Status of Hippo Components YAP and TAZ Using Phos-tag

  • Rui Chen
  • Steven W. Plouffe
  • Kun-Liang GuanEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1893)


Protein phosphorylation is one of the most important posttranslational modifications in cell signaling and regulation. Protein phosphorylation can be detected by site-specific phospho-antibodies, but generating these antibodies can be costly, time-consuming, and difficult. Recently, Phos-tag technology has been developed to detect protein phosphorylation. Here, we describe our method for using Phos-tag gels to compare the phosphorylation status of YAP and TAZ, the most important downstream effectors of the Hippo pathway.

Key words

Hippo pathway Phos-tag gel Phosphorylation YAP TAZ 



This work was supported by grants from the National Institutes of Health (CA196878, CA217642, and GM51586) to K.-L.G. S.W.P. is supported by the UCSD Graduate Training Program in Cellular and Molecular Pharmacology (T32 GM007752). K.-L.G. is a co-founder and has an equity interest in Vivace Therapeutics, Inc. The terms of this arrangement have been reviewed and approved by the University of California, San Diego, in accordance with its conflict of interest policies.


  1. 1.
    Yu FX, Zhao B, Guan KL (2015) Hippo pathway in organ size control, tissue homeostasis, and Cancer. Cell 163(4):811–828CrossRefGoogle Scholar
  2. 2.
    Zhao B et al (2010) A coordinated phosphorylation by Lats and CK1 regulates YAP stability through SCF beta-TRCP. Genes Dev 24(1):72–85CrossRefGoogle Scholar
  3. 3.
    Kinoshita E, Kinoshita-Kikuta E, Koike T (2009) Separation and detection of large phosphoproteins using Phos-tag SDS-PAGE. Nat Protoc 4(10):1513–1521CrossRefGoogle Scholar
  4. 4.
    Kinoshita-Kikuta E, Kinoshita E, Koike T (2009) Phos-tag beads as an immunoblotting enhancer for selective detection of phosphoproteins in cell lysates. Anal Biochem 389(1):83–85CrossRefGoogle Scholar
  5. 5.
    Plouffe SW et al (2016) Characterization of hippo pathway components by gene inactivation. Mol Cell 64(5):993–1008CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Pharmacology and Moores Cancer CenterUniversity of CaliforniaLa JollaUSA

Personalised recommendations