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Title: PRODUCTION AND CHARACTERIZATION OF MONOCLONAL ANTIBODIES DIRECTED AGAINST THE AH RECEPTOR

Author
item PERDEW, GARY - PURDUE UNIV
item ABBOTT, BARBARA - US ENVIRONMENTAL PROT AGY
item Stanker, Larry

Submitted to: Hybridoma
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 9/21/1995
Publication Date: N/A
Citation: N/A

Interpretive Summary: Dioxin is a highly toxic chemical that is found as an unwanted contaminant is various industrial processes. Exposure to dioxin has been shown to result in shrinkage of the thymus, a wasting syndrome, birth deformities, and tumor production. All of these effects are thought to be controlled by binding of the dioxin molecule to a receptor within the cell. This receptor ris referred to as the Ah receptor. In order to study this receptor and determine how the dioxin-Ah receptor complex result in such a diverse set of biological responses, it is necessary to have a probe for the receptor. In this paper we describe such a probe. The probe we developed is an antibody. Antibodies are proteins found in the blood that have the ability to bind to specific molecules. We engineered the production of antibodies that bind specifically with the dioxin Ah receptor. These antibody probes will have wide application as tools to study the effects of exposure to these highly toxic chemicals.

Technical Abstract: Six hybridomas secreting monoclonal antibodies that are specific for the N-terminal peptide sequence of the murine Ah receptor were isolated. These antibodies bind with high specificity to the Ah receptor on protein blots of Hepa 1c1c7 cytosol. Three IgG1 antibodies (Rpt 1,2,and 3) were capable of detecting 2 ng of receptor using peroxidase-goat anti-mouse IgG antibody yconjugate on a protein blot. Monoclonal antibody Rpt9 exhibited the greatest ability to immunoprecipitate the nondenatured 9S form of the Ah receptor and to visualize the AhR on liver tissue sections using immunohistochemical techniques. All of the monoclonal antibodies produced were able to bind to the mouse, rat, and human Ah receptor. These monoclonal antibodies should be useful in a wide number of applications in the study of Ah receptor biochemistry.