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Multiplex Immunofluorescence Staining Protocol for the Dual Imaging of Hypoxia-Inducible Factors 1 and 2 on Formalin-Fixed Paraffin-Embedded Samples

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Hypoxia

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2755))

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Abstract

Hypoxia is a common condition in rapidly proliferating tumors and occurs when oxygen delivery to the tissue is scarce. It is a prevalent feature in ~90% of solid tumors. The family of HIF (hypoxia-inducible factor) proteins—HIF1α and HIF2α—are the main transcription factors that regulate the response to hypoxia. These transcription factors regulate numerous downstream gene targets that promote the aggressiveness of tumors and therefore have been linked to worse prognosis in patients. This makes them a potential biomarker to be tested in the clinical setting to predict patient outcomes. However, HIFs have been notoriously challenging to immunolabel, in part due to their fast turnover under normal oxygen conditions. In this work, we developed a multiplexed immunofluorescence (mIF) staining protocol for the simultaneous detection of HIF1α and HIF2α in the same formalin-fixed paraffin-embedded (FFPE) tissue section.

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Acknowledgments

Work in the Gilkes lab has been supported by the National Cancer Institute, the Jayne Koskinas Ted Giovanis Foundation for Health and Policy, Cindy Rosencrans Fund for Metastatic Triple-Negative Breast Cancer, and the Emerson Collective.

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Authors and Affiliations

  1. Department of Oncology, The Sidney Kimmel Comprehensive Cancer Center, The Johns Hopkins University School of Medicine, Baltimore, MD, USA

    Harsh H. Oza & Daniele M. Gilkes

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  1. Harsh H. Oza
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  2. Daniele M. Gilkes
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Correspondence to Daniele M. Gilkes .

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  1. Department of Oncology Johns Hopkins, University School of Medicine, Baltimore, MD, USA

    Daniele M. Gilkes

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Oza, H.H., Gilkes, D.M. (2024). Multiplex Immunofluorescence Staining Protocol for the Dual Imaging of Hypoxia-Inducible Factors 1 and 2 on Formalin-Fixed Paraffin-Embedded Samples. In: Gilkes, D.M. (eds) Hypoxia. Methods in Molecular Biology, vol 2755. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3633-6_12

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  • DOI: https://doi.org/10.1007/978-1-0716-3633-6_12

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3632-9

  • Online ISBN: 978-1-0716-3633-6

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