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Nuclear factor-kappaB in middle ear epithelial cells: a methodological study using an ELISA.

Therese Ovesen·Thomas Q Barrett
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Abstract

OBJECTIVE: To contribute to the elucidation of the intracellular mechanisms relating to otitis media with effusion (OME) by detecting and quantifying nuclear factor (NF)-kappaB in middle ear epithelial cells by means of an ELISA. MATERIAL AND METHODS: Using an ELISA technique, NF-kappaB was identified and quantified in the cytoplasm of cultured epithelial cells from the rabbit middle ear exposed to a well-known inflammatory stimulus: lipopolysaccharide (LPS). RESULTS: NF-kappaB was measurable with the ELISA in a concentration range of 16-32 ng/microg total cell protein. LPS caused significant enhancement of NF-kappaB. Furthermore, the NF-kappaB concentration increased with the duration of LPS exposure. Western blot analysis confirmed the results obtained by ELISA. CONCLUSION: Further studies are required to determine the role of NF-kappaB in initiating and regulating the inflammatory response in the middle ear, including the significance of hyperoxia.

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AnimalsBlotting, WesternCells, CulturedEar, MiddleEnzyme-Linked Immunosorbent AssayEpithelial CellsNF-kappa BProbabilityRabbitsSensitivity and SpecificityComparative StudyJournal Article

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Therese Ovesen, Thomas Q Barrett. 2003. Nuclear factor-kappaB in middle ear epithelial cells: a methodological study using an ELISA.. https://doi.org/10.1080/00016480310001169

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