Development and characterization of polyclonal antibodies against the aryl hydrocarbon receptor protein family (AHR1, AHR2, and AHR repressor) of Atlantic killifish Fundulus heteroclitus
Merson, Rebeka R.
Franks, Diana G.
Karchner, Sibel I.
Hahn, Mark E.
MetadataShow full item record
KeywordAryl hydrocarbon receptor (AHR); AHR repressor (AHRR); Basic helix-loop-helix Per-ARNT-Sim (bHLH-PAS); Fish; Halogenated aromatic hydrocarbons (HAH);; Polynuclear aromatic hydrocarbons (PAH); Recombinant protein; Teleost
The aryl hydrocarbon receptor (AHR) and AHR repressor (AHRR) proteins regulate gene expression in response to some halogenated aromatic hydrocarbons and polycyclic aromatic hydrocarbons. The Atlantic killifish is a valuable model of the AHR signaling pathway, but antibodies are not available to fully characterize AHR and AHRR proteins. Using bacterially expressed AHRs, we developed specific and sensitive polyclonal antisera against the killifish AHR1, AHR2, and AHRR. In immunoblots, these antibodies recognized full-length killifish AHR and AHRR proteins synthesized in rabbit reticulocyte lysate, proteins expressed in mammalian cells transfected with killifish AHR and AHRR constructs, and AHR proteins in cytosol preparations from killifish tissues. Killifish AHR1 and AHR2 proteins were detected in brain, gill, kidney, heart, liver, and spleen. Antisera specifically precipitated their respective target proteins in immunoprecipitation experiments with in vitro-expressed proteins. Killifish ARNT2 co-precipitated with AHR1 and AHR2. These sensitive, specific, and versatile antibodies will be valuable to researchers investigating AHR signaling and other physiological processes involving AHR and AHRR proteins.
Author Posting. © The Authors, 2005. This is the author's version of the work. It is posted here by permission of Elsevier B. V. for personal use, not for redistribution. The definitive version was published in Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology 142 (2006): 85-94, doi:10.1016/j.cbpc.2005.10.013.
Suggested CitationPreprint: Merson, Rebeka R., Franks, Diana G., Karchner, Sibel I., Hahn, Mark E., "Development and characterization of polyclonal antibodies against the aryl hydrocarbon receptor protein family (AHR1, AHR2, and AHR repressor) of Atlantic killifish Fundulus heteroclitus", 2005-12-20, https://doi.org/10.1016/j.cbpc.2005.10.013, https://hdl.handle.net/1912/603
Showing items related by title, author, creator and subject.
Hestermann, Eli V. (Massachusetts Institute of Technology and Woods Hole Oceanographic Institution, 1999-12)Aryl hydrocarbon receptor (AHR) expression and activity was characterized in the teleost hepatoma cell line, PLHC-1. This work was carried out in order to gain insights into mechanisms of halogenated aromatic hydrocarbon ...
Involvement of Cytochrome P450 1A in the toxicity of aryl hydrocarbon receptor agonists : alteration arachidonic acid metabolism and production of reactive oxygen species Schlezinger, Jennifer J. (Massachusetts Institute of Technology and Woods Hole Oceanographic Institution, 1998-08)Two cytochrome P4501A-dependent mechanisms of aryl hydrocarbon receptor (AhR) agonist toxicity were examined in the marine teleost scup (Stenotomus chrysops), alteration of arachidonic acid (AA) metabolism and production ...
The molecular basis for differential dioxin sensitivity in birds : role of the aryl hydrocarbon receptor Karchner, Sibel I.; Franks, Diana G.; Kennedy, Sean W.; Hahn, Mark E. (National Academy of Sciences, 2006-04-10)2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) and related halogenated aromatic hydrocarbons (HAHs) are highly toxic to most vertebrate animals, but there are dramatic differences in sensitivity among species and strains. ...