Reverse Phase Protein Arrays—Quantitative Assessment of Multiple Biomarkers in Biopsies for Clinical Use

摘要: Reverse Phase Protein Arrays (RPPA) represent a very promising sensitive and precise high-throughput technology for the quantitative measurement of hundreds signaling proteins in biological clinical samples. This array format allows quantification one protein or phosphoprotein multiple samples under same experimental conditions at time. Moreover, it is suited signal transduction profiling small numbers cultured cells isolated from human biopsies, including formalin fixed paraffin embedded (FFPE) tissues. Owing to much easier sample preparation, as compared mass spectrometry based technologies, extraordinary sensitivity detection low-abundance over large linear range, RPPA have potential characterization deregulated interconnecting pathways networks limited amounts material routine settings. Current aspects technology, dilution curves, spotting, controls, detection, antibody validation, calculation levels are addressed.