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Quantification of serum Ovalbumin-specific immunoglobulin E titre via in vivo passive cutaneous anaphylaxis assay

Mincham, K., Leffler, J., Scott, N., Lauzon-Joset, J-F, Stumbles, P., Holt, P. and Strickland, D. (2019) Quantification of serum Ovalbumin-specific immunoglobulin E titre via in vivo passive cutaneous anaphylaxis assay. BIO-PROTOCOL, 9 (5).

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Murine models of allergic airway disease are frequently used as a tool to elucidate the cellular and molecular mechanisms of tissue-specific asthmatic disease pathogenesis. paramount to the success of these models is the induction of experimental antigen sensitization, as indicated by the presence of antigen-specific serum immunoglobulin E. The quantification of antigen-specific serum lgE is routinely performed via enzyme-linked immunosorbent assay. However, the reproducibility of these in vitro assays can vary dramatically in our experience. Furthermore, quantifying lgE via in vitro methodologies does not enable the functional relevance of circulating lgE levels to be considered. As a biologically appropriate alternative method, we describe herein a highly reproducible in vivo passive cutaneous anaphylaxis assay using Sprague Dawley rats for the quantification of ovalbumin-specific lgE in serum samples ovalbumin-sensitized murine models. Briefly, this in vivo assay involves subcutaneous injections of serum samples on the back of a Sprague Dawley rat, followed 24 h later by intravenous injection of ovalbumin and a blue detection dye. The subsequent result of antigen-lgE mediated inflammation and leakage of blue dye into the initial injection site indicates the presence of ovalbumin-specific lgE within the corresponding serum sample.

Item Type: Journal Article
Murdoch Affiliation(s): Medical, Molecular and Forensic Sciences
Publisher: Bio-Protocol
Copyright: © 2019 The Authors
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