Protein arrays III: Reverse-phase protein arrays

Yulin Yuan, Xia Hong, Zuan Tao Lin, Hongting Wang, Mikala Heon, Tianfu Wu

Research output: Chapter in Book/Report/Conference proceedingChapter

3 Scopus citations

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.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages279-289
Number of pages11
Volume1654
DOIs
StatePublished - 2017

Publication series

NameMethods in Molecular Biology
Volume1654
ISSN (Print)1064-3745

Keywords

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

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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