Development of an enzyme linked immunosorbent assay (ELISA) for the detection of Zebrafish Interleukin 1 beta upregulation Public Deposited

http://ir.library.oregonstate.edu/concern/honors_college_theses/6d570233g

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  • The study and utilization of Danio rerio (Zebrafish) as a model organism has increased greatly in recent decades. They are used in many areas of biological research, but one field that has used them extensively is immunology. Since zebrafish have been found to possess many orthologs to human genes, study of the relationship between gene transcription and protein production has been a key area of interest. One gene and protein of particular interest to the field of immunology is interleukin-1β (IL1β), a pro-inflammatory cytokine that is produced in an inactive pro-form before being post-translationally modified and excreted into its active form. While IL1β is a well-studied cytokine, research into its expression and regulation at the protein level is still growing. One reason for this is a lack of reagents and straightforward assays that can be used to detect changes in protein production with precision. The body of zebrafish research would benefit greatly from a sensitive assay based on monoclonal antibodies. In this paper, the generation and characterization of two monoclonal antibodies reactive against zebrafish IL-1β is reported. These were used in conjunction to develop an IL1β-specific sandwich ELISA assay system to detect native IL-1β produced by bacteria-stimulated zebrafish leucocytes and individually to develop a competitive ELISA for the same purpose.
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