REVERSE PHASE PROTEIN ARRAYS: FROM TECHNICAL AND ANALYTICAL FUNDAMENTALS TO APPLICATIONS

Espina V and Mueller C.

Reverse Phase Protein Arrays: From Technical and Analytical Fundamentals to Applications Singapore: Springer Singapore; 2019: 61-75.

doi: https://doi.org/10.1007/978-981-32-9755-5_4

Abstract

Reverse phase protein arrays (RPPA) are miniature dot blots constructed using robotic arrayers to deposit protein containing samples onto nitrocellulose-coated glass slides. Reverse phase protein arrays address the challenge of quantifying low-abundance proteins and posttranslationally modified proteins in cellular lysates and body fluids. RPPA technology is ideally suited to biomarker discovery, signal pathway profiling, functional phenotype analysis, and mechanism of action studies for drug discovery. Each array is fabricated with specimens, controls, and calibrators, thus providing a complete assay on each slide. Constructing a reverse phase protein array initially consists of selecting an arrayer, pin type, print head configuration, and nitrocellulose slide that is optimized for the particular specimen type and protein detection method. Herein we present the nuances of RPPA fabrication and study design using a solid pin arrayer and nitrocellulose-coated slides.