lifesciences
Innopsys

Reverse Phase Protein Arrays for Cell Signaling: Using the InnoScan 710-IR scanner improves sensitivity and dynamic range.

Reverse Phase Protein Arrays (RPPAs) are increasingly used to study cell signaling as they are highly multiplexed and allow precise quantification of protein of interests in biological samples. Such arrays promote automation, simplified work flow, and preserve sample. One of the principal challenges faced by RPPAs is to find the best combination of an array surface that will support high binding levels of viable protein probes, and a detection method that will provide high sensitivity and dynamic range, especially as lysates are complex samples. Fluorescence detection provides greater sensitivity and dynamic range, but is severely limited for use with nitrocellulose arrays because nitrocellulose has strong auto-fluorescence in the classic 532nm or 635nm wavelengths. A solution to this problem is to scan in the infrared wavelengths which allows diminishing background noise. With the new InnoScan 710-IR scanner it is now possible to combine infrared detection with high sensitivity and quality images. Here we prove that the InnoScan 710-IR improves the detected signal by a factor of up to 2.7 especially for low concentrations of arrayed cell lysates and extends the dynamic range of detection compared to the existing infrared scanner.

 

Marie-Laure Schneider1, Adriana Lagraulet2, Richard Pasquesi3 and Nathan Moerke4

  • 1,3 Innopsys Inc. Chicago, IL
  • 2 Innopsys, Parc Activestre 31390 Carbonne, France
  • 4 Laboratory of Peter Sorger, Department of Systems Biology, Harvard Medical School, Boston, MA

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