Open Access

Development of multiplexed protein profiling and detection using near infrared detection of reverse-phase protein microarrays

  • Valerie S. Calvert1,
  • Yihui Tang2,
  • Vince Boveia2,
  • Julie Wulfkuhle3,
  • Amy Schutz-Geschwender2,
  • D. Michael Olive2,
  • Lance A. Liotta3 and
  • Emanuel F. PetricoinIII1Email author
Clinical Proteomics1:11081

DOI: 10.1385/CP:1:1:081

Abstract

Protein microarrays have been recently employed for signal pathway profiling and high-throughput protein expression analysis. Reversephase arrays, where the array consists of immobilized analytes and lysates has especially shown promise in low abundance analyte detection and signal pathway profiling using phospho-specific antibodies. A limitation to current reverse phase array methodology is the inability to multiplex proteomic-based endpoints as each array can only report one analyte endpoint. In this study, we report on the use of a dual dye based approach that can effectively double the number of endpoints observed per array allowing, for example, both phosphospecific and total protein levels to be measured and analyzed at once. The method utilizes antibody bound dyes that emit in the infrared spectral region as a means of sensitive and specific detection.

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