Improved Immunohistochemical Method for Detecting Hypoxia Gradients in Mouse Tissues and Tumors | Zendy

Michael Samoszuk | Zendy; James Walter | Zendy; Eugene Mechetner | Zendy

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Improved Immunohistochemical Method for Detecting Hypoxia Gradients in Mouse Tissues and Tumors

Author(s) -

Michael Samoszuk,

James Walter,

Eugene Mechetner

Publication year - 2004

Publication title -

journal of histochemistry and cytochemistry

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.971

H-Index - 124

eISSN - 1551-5044

pISSN - 0022-1554

DOI - 10.1369/jhc.4b6248.2004

Subject(s) - immunohistochemistry , pathology , horseradish peroxidase , monoclonal antibody , antibody , primary and secondary antibodies , hypoxia (environmental) , staining , rodent , antigen , biology , kidney , microbiology and biotechnology , chemistry , immunology , medicine , biochemistry , enzyme , oxygen , endocrinology , ecology , organic chemistry

We describe an improved immunohistochemical procedure for detecting regions of hypoxia in normal organs and tumors in mice. The method employs a primary fluorescein-conjugated mouse monoclonal antibody directed against pimonidazole protein adducts that are created in hypoxic tissues and a secondary mouse anti-fluorescein antibody that is conjugated to horseradish peroxidase. Using these reagents, we clearly visualized the regions of relative hypoxia in implanted tumors in mice as well as in normal organs such as liver and kidney. Significantly, the resulting tissue sections were remarkably free of the background staining that is characteristically observed when rodent antibodies are used to detect antigens in rodent tissues.

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