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

Methods Mol Biol. 2018:1709:307-320. doi: 10.1007/978-1-4939-7477-1_22.

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).

Keywords: Flow cytometry; Formalin fixed paraffin-embedded (FFPE) sections; Glioblastoma multiforme; Hsp70 antibody epitope; Immunohistochemistry; Intracellular and membrane-bound heat shock protein 70 (Hsp70).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aged
  • Antibodies, Monoclonal*
  • Biomarkers, Tumor / analysis
  • Cell Line, Tumor
  • Cell Membrane / metabolism
  • Cytoplasm / metabolism
  • Cytosol / metabolism
  • Female
  • Flow Cytometry / methods*
  • Glioblastoma / metabolism*
  • HSP70 Heat-Shock Proteins / analysis*
  • Humans
  • Immunohistochemistry / methods*
  • Male
  • Middle Aged

Substances

  • Antibodies, Monoclonal
  • Biomarkers, Tumor
  • HSP70 Heat-Shock Proteins