Preparation and use of reverse protein microarrays

Elisa Pin, Giulia Federici, Emanuel F. Petricoin

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Reverse-phase protein array (RPPA) is a multiplex, high-throughput proteomic technique for profiling the activation status of signal transduction pathways involved in cancer survival and progression, potentially allowing for identification of new biomarkers and drug targets. On RPPA, the entire patient proteome is immobilized on a spot and single proteins can be quantified across a set of samples, spotted on the same array, with high specificity and sensitivity. Array immunostaining and signal amplification systems are used to generate a signal proportional to the concentration of the analyte. Dedicated scanners and software are used to detect spots, measure intensity, subtract background, normalize signal, and generate a numeric value as output. The generated output file is then analyzed using several different bioinformatic and biostatistical tools. In this unit, the RPPA procedure is described in depth, from sample handling and preparation to data analysis, with particular emphasis on tissue sample analysis.

Original languageEnglish
Pages (from-to)27.7.1-27.29
JournalCurrent Protocols in Protein Science
Volume2014
DOIs
Publication statusPublished - 2014

Fingerprint

Protein Array Analysis
Microarrays
Proteins
Proteome
Computational Biology
Proteomics
Signal transduction
Signal Transduction
Biomarkers
Software
Bioinformatics
Amplification
Sensitivity and Specificity
Survival
Chemical activation
Throughput
Tissue
Pharmaceutical Preparations
Neoplasms

Keywords

  • Analysis
  • Antibody
  • Cells
  • Microarray
  • Protein
  • Tissues

ASJC Scopus subject areas

  • Biochemistry
  • Structural Biology

Cite this

Preparation and use of reverse protein microarrays. / Pin, Elisa; Federici, Giulia; Petricoin, Emanuel F.

In: Current Protocols in Protein Science, Vol. 2014, 2014, p. 27.7.1-27.29.

Research output: Contribution to journalArticle

Pin, Elisa ; Federici, Giulia ; Petricoin, Emanuel F. / Preparation and use of reverse protein microarrays. In: Current Protocols in Protein Science. 2014 ; Vol. 2014. pp. 27.7.1-27.29.
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