RPPA Protein Profiling Services

Reverse Phase Protein Arrays (RPPAs) are planar arrays of printed cell and tissue lysates. Using RPPAs, changes of protein levels and their activation status (e.g. phospho-signaling) can be detected in numerous cell and tissue sources, e.g. cancer. The miniaturized array and direct immunoassay format (based on the Zeptosens platform) facilitate efficient and comprehensive analyses of cellular signaling networks for hundreds of proteins and samples in parallel with low sample consumption.

New! Through careful testing via DigiWest and Western blotting, we have significantly expanded our core repertoire of well-validated RPPA antibodies. The result is a list of 500+ antibodies, which you can now use to fully customize your list of analytes. Or you simply rely on 50+ pre-defined pathway panels. Just contact us to get the list.

Key features

  • Established technology, proven to add value
  • ISO9001 compliant, robust Standard Operating Procedures (SOPs)
  • More than 10 years of experience in designing and performing RPPA studies for life science industry and academic research
  • For studies of hundreds of samples with dozens to hundreds of protein markers
  • Select analytes from our list of 500+ pre-validated pathway marker antibodies for total proteins and post-translationally modified forms
  • Additional antibodies for signaling proteins of interest can be validated upon request

Established applications

  • Lead compound mechanism-of-action and translational studies
  • Quantification of on/off target & downstream signaling effects
  • Transient selective pathway inhibition and cross-pathway effects
  • Biomarker screening/verification of preclinical & clinical samples
  • Mechanisms and markers for treatment efficacy and resistance
  • Providing rationale for treatment combination strategies to overcome resistance mechanisms

Typical samples

  • Cell cultures (2D; 3D, e.g. spheroids, microtissues)
  • Animal tissues (different organs)
  • Xenografts, PDX tumor models
  • Patient tissues, tumor biopsies, either fresh frozen (FF) or formalin-fixed paraffin-embedded (FFPE)
  • Minimum sample amount required for a typical RPPA study: 50 µg total protein at 3 µg/µl protein concentration

References

  • Pawlak M, Carragher NO. Reverse Phase Protein Arrays Elucidate Mechanisms-of-action and Phenotypic Response in 2D and 3D models. Drug Discov Today Technol 2017 | Contact an author.

  • Bader S, Zajac M, Friess T, Ruge E, Rieder N, Gierke B, Heubach Y, Thomas M, Pawlak M. Evaluation of Protein Profiles From Treated Xenograft Tumor Models Identifies an Antibody Panel for FFPE Tissue Analysis by RPPA. Mol Cell Proteomics 2015 | Contact an author.

  • Tegnebratt T, Ruge E, Bader S, Ishii N, Aida S, Yoshimura Y, Ooi CH, Lu L, Mitsios N, Meresse V, Mulder J, Pawlak M, Venturi M, Tessier J, Stone-Elander S. Evaluation of Efficacy of a New MEK Inhibitor, RO4987655, in Human Tumor Xenografts by [(18)F] FDG-PET Imaging Combined with Proteomic Approaches. EJNMMI Research 2014 | Contact an author.

  • Akbani R, Becker KF, Carragher N, Goldstein T, de Koning L, Korf U, Liotta L, Mills GB, Nishizuka SS, Pawlak M, Petricoin EF 3rd, Pollard HB, Serrels B, Zhu J. Realizing the Promise of RPPA for Clinical, Translational, and Basic Research: a Workshop Report: the RPPA Society. Mol Cell Proteomics 2014 | Contact an author.

  • Assadi M, Lamerz J, Jarutat T, Farfsing A, Paul H, Gierke B, Breitinger E, Templin MF, Essioux L, Arbogast S, Venturi M, Pawlak M, Langen H, Schindler T. Multiple Protein Analysis of FFPE Tissue Samples with RPPA. Mol Cell Proteomics 2013 | Contact an author.

  • Pirnia F, Pawlak M, Thallinger GG, Gierke B, Templin MF, Kappeler A, Betticher DC, Gloor B, Borner MM. Novel Functional Profiling Approach Combining RPPA and Human 3-D ex vivo Tissue Cultures: Expression of Apoptosis-related Proteins in Human Colon Cancer. Proteomics 2009 | Contact an author.

  • Saturno G, Pesenti M, Cavazzoli C, Rossi A, Giusti AM, Gierke B, Pawlak M, Venturi M. Expression of Serine/Threonine Protein-kinases and Related Factors in Normal Monkey and Human Retinas: the Mechanistic Understanding of a CDK2 Inhibitor Induced Retinal Toxicity. Toxicologic Pathology 2007 | Contact an author.

  • Pawlak M, Schick E, Bopp MA, Schneider MJ, Oroszlan P, Ehrat M. Zeptosens Protein Microarrays: A Novel High-Performance Microarray Platform for Low Abundance Protein Analysis. Proteomics 2002 | Contact an author.