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Murine fashions of allergic airway illness are regularly used as a device to elucidate the mobile and molecular mechanisms of tissue-specific asthmatic illness pathogenesis. Paramount to the success of these fashions is the induction of experimental antigen sensitization, as indicated by the presence of antigen-specific serum immunoglobulin E. The quantification of antigen-specific serum IgE is routinely carried out by way of enzyme-linked immunosorbent assay.
However, the reproducibility of these in vitro assays can range dramatically in our expertise. Furthermore, quantifying IgE by way of in vitro methodologies doesn’t allow the practical relevance of circulating IgE ranges to be thought-about. As a biologically acceptable various technique, we describe herein a extremely reproducible in vivo passive cutaneous anaphylaxis assay utilizing Sprague Dawley rats for the quantification of ovalbumin-specific IgE in serum samples from ovalbumin-sensitized murine fashions.
Briefly, this in vivo assay includes subcutaneous injections of serum samples on the again of a Sprague Dawley rat, adopted 24 h later by intravenous injection of ovalbumin and a blue detection dye. The subsequent consequence of antigen-IgE mediated irritation and leakage of blue dye into the preliminary injection web site signifies the presence of ovalbumin-specific IgE throughout the corresponding serum pattern.