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

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...

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Bibliographic Details
Published in:The journal of histochemistry and cytochemistry 2004-06, Vol.52 (6), p.837-839
Main Authors: Samoszuk, Michael K, Walter, James, Mechetner, Eugene
Format: Article
Language:English
Subjects:
Animals
Antibodies, Monoclonal
Cell Hypoxia
Fluorescein-5-isothiocyanate - chemistry
Fluorescent Dyes - chemistry
Immunohistochemistry - methods
Mice
Neoplasms - metabolism
Nitroimidazoles - chemistry
Nitroimidazoles - immunology
Organ Specificity
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Description
Summary: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.
ISSN:0022-1554
1551-5044
DOI:10.1369/jhc.4B6248.2004