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Chaperones pp 307–320Cite as

Immunohistochemical and Flow Cytometric Analysis of Intracellular and Membrane-Bound Hsp70, as a Putative Biomarker of Glioblastoma Multiforme, Using the cmHsp70.1 Monoclonal Antibody

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1709))

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. 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 and membrane-bound Hsp70 as a potential tumor biomarker in glioblastoma multiforme, herein, we describe protocols for the staining of cytosolic Hsp70 in tumor formalin fixed paraffin-embedded (FFPE) sections using immunohistochemistry, and for plasma membrane-bound Hsp70 by multi-parametric flow cytometry using the cmHsp70.1 monoclonal antibody (mAb).

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Acknowledgments

Gabriele Multhoff is supported by grants of the Deutsche Forschungsgemeinschaft (SFB824/2; DFG INST95/980-1 FUGG, INST411/37-1 FUGG), the DFG Cluster of Excellence: Munich-Centre for Advanced Photonics (MAP), the Bundesministerium für Forschung und Technologie (BMBF Innovative Therapies, 01GU0823; Kompetenzverbund Strahlenforschung 02NUK038A), 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 from the John and Lucille van Geest Foundation, Nottingham Trent University, the European Regional Development Fund (ERDF) via the Healthcare and Bioscience iNET, as delivered by Medilink East Midlands, UK and the Headcase Cancer Trust.

The University of Portsmouth Brain Tumour Research Centre is core funded by Brain Tumour Research.

Author information

Authors and Affiliations

  1. Department of Radiation Oncology, Klinikum Rechts der Isar, Technische Universität München, Ismaninger Str. 22, Munich, 81675, Germany

    Stefan Stangl, Helena Fellinger, A. Graham Pockley & Gabriele Multhoff

  2. John van Geest Cancer Research Centre, School of Science and Technology, Nottingham Trent University, Clifton Campus, Nottingham, NG11 8NS, UK

    Gemma A. Foulds & A. Graham Pockley

  3. Brain Tumour Research Centre, Institute of Biomedical and Biomolecular Sciences, School of Pharmacy and Biomedical Sciences, University of Portsmouth, Portsmouth, PO1 2DT, UK

    Geoffrey J. Pilkington

  4. CCG-Innate Immunity in Tumor Biology, Helmholtz Zentrum München, Ingolstädter Landstrasse 1, Neuherberg, 85764, Germany

    Gabriele Multhoff

Authors
  1. Stefan Stangl
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  2. Gemma A. Foulds
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  3. Helena Fellinger
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  4. Geoffrey J. Pilkington
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  5. A. Graham Pockley
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  6. Gabriele Multhoff
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Corresponding author

Correspondence to Gabriele Multhoff .

Editor information

Editors and Affiliations

  1. Department of Radiation, Oncology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts, USA

    Stuart K. Calderwood

  2. Department of Urology, Weis Center for Research, Geisinger Clinic, Danville, Pennsylvania, USA

    Thomas L. Prince

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Stangl, S., Foulds, G.A., Fellinger, H., Pilkington, G.J., Pockley, A.G., Multhoff, G. (2018). Immunohistochemical and Flow Cytometric Analysis of Intracellular and Membrane-Bound Hsp70, as a Putative Biomarker of Glioblastoma Multiforme, Using the cmHsp70.1 Monoclonal Antibody. In: Calderwood, S., Prince, T. (eds) Chaperones. Methods in Molecular Biology, vol 1709. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7477-1_22

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  • DOI: https://doi.org/10.1007/978-1-4939-7477-1_22

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7476-4

  • Online ISBN: 978-1-4939-7477-1

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