70kDa的主要应激诱导（应激）蛋白70（Hsp70）在高度侵袭性肿瘤细胞中经常过度表达，因此可能作为侵略性疾病和/或治疗耐药性的肿瘤特异性生物标志物。我们先前证明，与正常细胞相反，肿瘤细胞在其细胞膜上表现出Hsp70。为了阐明细胞内、与细胞膜结合以及细胞外Hsp70作为癌症潜在肿瘤标志物的作用，本文描述了用免疫组织化学染色法对脑胶质母细胞瘤患者的肿瘤福尔马林固定石蜡包埋（FFPE）切片中的细胞质Hsp70进行染色的协议，用多参数流式细胞术（cmHsp70.1单克隆抗体）检测由多种癌细胞衍生的细胞膜结合Hsp70的表达，并使用独特的酶联免疫吸附测定（ELISA）在癌症患者循环中测量自由和囊泡相关的Hsp70。© 2023. 作者，在Springer Science+Business Media, LLC的独家许可下，属于Springer Nature的一部分。

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).© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.