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Validation of Antibodies Used to Study Hypoxia Inducible Factors in Two Species of Fundulus

Hypoxia inducible factors (HIFs) are transcription factors and the master regulators of oxygen-dependent gene expression in animals. The focus of this thesis is the distribution of HIF protein in tissues of the fish Fundulus heteroclitus and F. grandis, two widespread species that occur in naturally hypoxic waters. Polyclonal antibodies against HIF-1α, HIF-2α, and HIF-3α were tested on proteins made in vitro and on extracts made from several tissues of normoxic and hypoxic fish. Antibodies against HIF-1α and 3α bound specifically to full length protein made in vitro, and produced bands on western blots of nuclear extracts of near the expected molecular weights for these proteins. Hypoxic exposure did not markedly increase the intensity of these bands, and mass spectrometry failed to identify HIF-1α and 3α peptides in excised gel bands. Thus, further tests of antibody specificity are needed before the tissue distribution of HIF in these fish can be confidently assessed.

Identiferoai:union.ndltd.org:uno.edu/oai:scholarworks.uno.edu:td-2693
Date17 May 2013
CreatorsHill, Jenna D.
PublisherScholarWorks@UNO
Source SetsUniversity of New Orleans
Detected LanguageEnglish
Typetext
Formatapplication/pdf
SourceUniversity of New Orleans Theses and Dissertations

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