Lysate Preparation for Reverse Phase Protein Arrays

Protein lysates from a variety of sample materials, e.g., cell lines, serum, frozen tissues, formalin-fixed and paraffin-embedded (FFPE) tissues, and fine needle aspirates, are compatible with the analysis using reverse phase protein arrays (RPPAs). Due to this diversity of input material, lysate preparation is one of the critical steps for obtaining reliable RPPA results. The challenges include, but are not limited to, achieving complete solubilization, avoiding chemical or enzymatic protein modifications, and degradation. In this chapter, preparations of lysates for RPPA analysis are described, focusing on human tissue samples. Special emphasis is given to recently published ISO International Standards for the preanalytical phase.