Abstract
Allergen-specific Immunoglobulin E (IgE) determination lies at the heart of diagnosis of sensitization to food and other allergens. In the past few years, reporter systems capable of detecting the presence of allergen-specific IgE have been developed by several labs. These rely on humanized rat basophil leukemia cell lines stably transfected with reporter genes such as firefly luciferase. In this chapter, we describe protocols for the use of the RS-ATL8 cell line (IgE cross-linking-induced luciferase expression; EXiLE) in 96-well and 384-well formats. We also describe optional treatment steps for enveloped virus and complement inactivation.
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Acknowledgments
E.A.A. was funded by a University of Nottingham Vice-Chancellor’s Scholarship for Research Excellence (International) and the Gordon Memorial College Trust Fund. Sera from milk allergic individuals and fractionated milk allergens were kindly provided by M. Alcocer, University of Nottingham. Our thanks to Colin Fitzsimmons (University of Cambridge), who introduced us to the use of the resin for detergent removal.
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Ali, E.A., Nakamura, R., Falcone, F.H. (2017). Use of Humanized RS-ATL8 Reporter System for Detection of Allergen-Specific IgE Sensitization in Human Food Allergy. In: Lin, J., Alcocer, M. (eds) Food Allergens. Methods in Molecular Biology, vol 1592. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6925-8_12
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DOI: https://doi.org/10.1007/978-1-4939-6925-8_12
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