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Related Experiment Video

Updated: Mar 15, 2026

The Use of Reverse Phase Protein Arrays RPPA to Explore Protein Expression Variation within Individual Renal Cell Cancers
12:22

The Use of Reverse Phase Protein Arrays RPPA to Explore Protein Expression Variation within Individual Renal Cell Cancers

Published on: January 22, 2013

34.2K

Reverse Phase Protein Arrays for Compound Profiling.

Nathan Moerke1, Mohammad Fallahi-Sichani2

  • 1Harvard Medical School-ICCB-Longwood Screening Facility, Boston, Massachusetts.

Current Protocols in Chemical Biology
|September 14, 2016
PubMed
Summary
This summary is machine-generated.

Reverse phase protein arrays (RPPAs) enable high-throughput analysis of protein and post-translational modification (PTM) levels in cells and tissues. This method details RPPA generation and analysis for cell signaling studies.

Keywords:
BRAFantibodiescell signalingfluorescencemelanomareverse phase protein arrays

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Area of Science:

  • Biochemistry and Molecular Biology
  • Proteomics
  • Cellular Signaling

Background:

  • Reverse phase protein arrays (RPPAs) are powerful tools for quantifying protein and post-translational modification (PTM) levels.
  • Existing methods require detailed protocols for sample preparation and array analysis.

Purpose of the Study:

  • To describe a comprehensive procedure for generating and analyzing Reverse Phase Protein Arrays (RPPAs).
  • To provide adaptable methods for studying cell signaling pathways in response to various perturbations.

Main Methods:

  • Immobilization of cell or tissue lysates onto solid supports.
  • Probing with specific primary antibodies against target proteins or PTMs.
  • Detailed procedures for cell treatment, lysate preparation, RPPA generation, probing, imaging, and data analysis.

Main Results:

  • Established a robust and adaptable protocol for RPPA creation and analysis.
  • Demonstrated the utility of RPPAs for large-scale, high-throughput protein and PTM level measurements.
  • Highlighted the affordability and multiplexing capabilities of the RPPA technique.

Conclusions:

  • The described RPPA procedures are suitable for extensive studies of cell signaling.
  • RPPAs offer a cost-effective and highly multiplexable approach for proteomic analysis.
  • The methods are readily adaptable for investigating cellular responses to drugs and other stimuli.