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Protein Arrays III: Reverse-Phase Protein Arrays

  • Yulin Yuan
  • Xia Hong
  • Zuan-Tao Lin
  • Hongting Wang
  • Mikala Heon
  • Tianfu WuEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1654)

Abstract

The reverse-phase protein array (RPPA) is to use highly specific antibodies to interrogate pan or posttranslationally modified protein targets, such as phosphorylated proteins, particularly the proteins involved in cell signaling pathways. In this protocol we will cover the preparation of cell (or tissue) lysates, sample printing, antibody validation, antibody interrogation, signal amplification steps, imaging and data analysis. In this protocol, colorimetric catalyzed signal amplification (CSA) chemistry, fluorescence and near-infrared (NIR) based detection methods will be described.

Key words

Reverse-phase protein array Antibody validation Cell signaling pathways Near-infrared (NIR) fluorescence 

Notes

Acknowledgments

This work was partly supported by a grant from the Lupus Research Institute to T.W. and a startup fund from the University of Houston to T.W.

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Yulin Yuan
    • 1
    • 2
  • Xia Hong
    • 1
    • 3
  • Zuan-Tao Lin
    • 1
  • Hongting Wang
    • 1
    • 4
  • Mikala Heon
    • 1
  • Tianfu Wu
    • 1
    Email author
  1. 1.Department of Biomedical EngineeringUniversity of HoustonHoustonUSA
  2. 2.Department of Clinical LaboratoryThe People’s Hospital of Guangxi Zhuang Autonomous RegionNanningChina
  3. 3.Department of NursingFujian Health CollegeFuzhouChina
  4. 4.National Pharmacology Laboratory of Chinese MedicineBasic Medical College, Wannan Medical CollegeWuhuChina

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