Staining Against Phospho-H2AX (γ-H2AX) as a Marker for DNA Damage and Genomic Instability in Cancer Tissues and Cells

Anika Nagelkerke; Paul N Span

doi:10.1007/978-3-319-26666-4_1

Staining Against Phospho-H2AX (γ-H2AX) as a Marker for DNA Damage and Genomic Instability in Cancer Tissues and Cells

Anika Nagelkerke, Paul N Span

Research output: Contribution to journal › Article › Academic › peer-review

49 Citations (Scopus)

Abstract

Phospho-H2AX or γ-H2AX- is a marker of DNA double-stranded breaks and can therefore be used to monitor DNA repair after, for example, irradiation. In addition, positive staining for phospho-H2AX may indicate genomic instability and telomere dysfunction in tumour cells and tissues. Here, we provide a protocol to perform immunostaining for phospho-H2AX on cells, cryosections and formalin-fixed, paraffin-embedded tissues. Crucial steps in the protocol and troubleshooting suggestions are indicated. We also provide suggestions on how to combine staining against γ-H2AX with stainings against components of the tumour microenvironment, such as hypoxia and blood vessels.

Original language	English
Pages (from-to)	1-10
Number of pages	10
Journal	Advances in experimental medicine and biology
Volume	899
DOIs	https://doi.org/10.1007/978-3-319-26666-4_1
Publication status	Published - 22-Jun-2016
Externally published	Yes

Keywords

Biomarkers/metabolism
Cryoultramicrotomy
DNA Damage
Genomic Instability
Histones/metabolism
Humans
Neoplasms/genetics
Paraffin Embedding
Phosphorylation
Staining and Labeling/methods
Tissue Fixation

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10.1007/978-3-319-26666-4_1

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Cite this

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title = "Staining Against Phospho-H2AX (γ-H2AX) as a Marker for DNA Damage and Genomic Instability in Cancer Tissues and Cells",

abstract = "Phospho-H2AX or γ-H2AX- is a marker of DNA double-stranded breaks and can therefore be used to monitor DNA repair after, for example, irradiation. In addition, positive staining for phospho-H2AX may indicate genomic instability and telomere dysfunction in tumour cells and tissues. Here, we provide a protocol to perform immunostaining for phospho-H2AX on cells, cryosections and formalin-fixed, paraffin-embedded tissues. Crucial steps in the protocol and troubleshooting suggestions are indicated. We also provide suggestions on how to combine staining against γ-H2AX with stainings against components of the tumour microenvironment, such as hypoxia and blood vessels. ",

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AU - Span, Paul N

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N2 - Phospho-H2AX or γ-H2AX- is a marker of DNA double-stranded breaks and can therefore be used to monitor DNA repair after, for example, irradiation. In addition, positive staining for phospho-H2AX may indicate genomic instability and telomere dysfunction in tumour cells and tissues. Here, we provide a protocol to perform immunostaining for phospho-H2AX on cells, cryosections and formalin-fixed, paraffin-embedded tissues. Crucial steps in the protocol and troubleshooting suggestions are indicated. We also provide suggestions on how to combine staining against γ-H2AX with stainings against components of the tumour microenvironment, such as hypoxia and blood vessels.

AB - Phospho-H2AX or γ-H2AX- is a marker of DNA double-stranded breaks and can therefore be used to monitor DNA repair after, for example, irradiation. In addition, positive staining for phospho-H2AX may indicate genomic instability and telomere dysfunction in tumour cells and tissues. Here, we provide a protocol to perform immunostaining for phospho-H2AX on cells, cryosections and formalin-fixed, paraffin-embedded tissues. Crucial steps in the protocol and troubleshooting suggestions are indicated. We also provide suggestions on how to combine staining against γ-H2AX with stainings against components of the tumour microenvironment, such as hypoxia and blood vessels.

KW - Biomarkers/metabolism

KW - Cryoultramicrotomy

KW - DNA Damage

KW - Genomic Instability

KW - Histones/metabolism

KW - Humans

KW - Neoplasms/genetics

KW - Paraffin Embedding

KW - Phosphorylation

KW - Staining and Labeling/methods

KW - Tissue Fixation

U2 - 10.1007/978-3-319-26666-4_1

DO - 10.1007/978-3-319-26666-4_1

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