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Immunohistochemical, Flow Cytometric, and ELISA-Based Analyses of Intracellular, Membrane-Expressed, and Extracellular Hsp70 as Cancer Biomarkers

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Chaperones

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2693))

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Abstract

The major stress-inducible 70 kDa heat shock (stress) protein 70 (Hsp70) is frequently overexpressed in highly aggressive tumor cells and thus might serve as a tumor-specific biomarker of aggressive disease and/or therapeutic resistance. We have previously shown that, in contrast to normal cells, tumor cells present Hsp70 on their plasma membrane. In order to elucidate the role of intracellular, membrane-bound and extracellular Hsp70 as a potential tumor biomarker in cancer, herein we describe protocols for the staining of cytosolic Hsp70 in tumor formalin-fixed paraffin-embedded (FFPE) sections from patients with glioblastoma multiforme using immunohistochemistry, for detecting the expression of plasma membrane-bound Hsp70 by a range of cancer-derived cells using multi-parametric flow cytometry using the cmHsp70.1 monoclonal antibody (mAb) and for the measurement of free and vesicular-associated Hsp70 in the circulation of patients with cancer using a unique enzyme-linked immunosorbent assay (ELISA).

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Acknowledgments

Gabriele Multhoff has received funding from the Deutsche Forschungsgemeinschaft (SFB824/2; DFG INST95/980-1 FUGG, INST411/37-1 FUGG, DFG KU3500/2-1, DFG STA1520/1-1), the DFG Cluster of Excellence: Munich-Centre for Advanced Photonics (MAP), the Bundesministerium für Forschung und Technologie (BMBF Innovative Therapies, 01GU0823; Kompetenzverbund Strahlenforschung 02NUK038A), the Bundesministerium für Wirtschaft und Energie BMWi (ZF4320104AJ8, ZF4320102CS7) EU-CELLEUROPE (315963), and the German Cancer Consortium Radiation Oncology Group, Munich (DKTK-ROG). The John van Geest Cancer Research Centre is supported by funding received from the John and Lucille van Geest Foundation and the Headcase Cancer Trust. GJP was awarded grant funding from the Ollie Young Foundation and Brain Tumour Research.

Conflict of Interest Statement

AGP is CEO of multimmune GmbH; GM is the founder, former CEO, and current CSO of multimmune GmbH.

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Authors and Affiliations

  1. Radiation Immuno-Oncology Group, Center for Translational Cancer Research Technische Universität München (TranslaTUM), Department of Radiation Oncology, Klinikum rechts der Isar, Technische Universität München (TUM), Munich, Germany

    Ali Bashiri Dezfouli, Stefan Stangl, Philipp Lennartz & Gabriele Multhoff

  2. John van Geest Cancer Research Centre, School of Science and Technology, Nottingham Trent University, Nottingham, UK

    Gemma A. Foulds & A. Graham Pockley

  3. Brain Tumour Research Centre, School of Pharmacy and Biomedical Sciences, Institute of Biomedical and Biomolecular Sciences, University of Portsmouth, Portsmouth, UK

    Geoffrey J. Pilkington

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  1. Ali Bashiri Dezfouli
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  2. Stefan Stangl
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  3. Gemma A. Foulds
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  4. Philipp Lennartz
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  5. Geoffrey J. Pilkington
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  6. A. Graham Pockley
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  7. Gabriele Multhoff
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Correspondence to Gabriele Multhoff .

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Editors and Affiliations

  1. Harvard Medical School, Beth Israel Deaconess Medical Center, Boston, MA, USA

    Stuart K. Calderwood

  2. Danville, PA, USA

    Thomas L. Prince

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Dezfouli, A.B. et al. (2023). Immunohistochemical, Flow Cytometric, and ELISA-Based Analyses of Intracellular, Membrane-Expressed, and Extracellular Hsp70 as Cancer Biomarkers. In: Calderwood, S.K., Prince, T.L. (eds) Chaperones. Methods in Molecular Biology, vol 2693. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3342-7_23

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  • DOI: https://doi.org/10.1007/978-1-0716-3342-7_23

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  • Publisher Name: Humana, New York, NY

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