Hrp conjugated goat anti mouse igg2c

Sera were collected from naïve or Trichuris muris infected mice at 19 days post-infection and stored at −30°C for side-by-side analysis. Total circulating IgE concentration was determined using the Mouse IgE ELISA Set (BD Biosciences) by following the manufacturer’s protocol. Serially diluted sera at 1:5, 1:25, and 1:125 were used for IgE detection. Trichuris antigen-specific IgG1 and IgG2c concentrations were measured via antigen-specific ELISA. Briefly, Immulon 4HBX Extra High Binding plates (Thermo Scientific) were coated with E/S Trichuris antigen in ELISA coating buffer (PBS supplemented with 0.1 M sodium carbonate (Sigma-Aldrich), 0.1M sodium bicarbonate (Sigma-Aldrich), and 1mM sodium Azide (Sigma-Aldrich) at pH 9.6). Sera were serially diluted two-fold from 1:20 to 1:2560 and incubated in the antigen-coated Immulon 4HBX plates. Antigen-specific IgG1 or IgG2c bound to the immobilized Trichuris antigen was then detected with HRP-conjugated goat anti-mouse IgG1 (Thermo Scientific) or HRP-conjugated goat anti-mouse IgG2c (Thermo Scientific). For IgE, the plates were developed with TMB substrate (Thermo Scientific) and KPL TMB Stop Solution (SeraCare), and optical density (OD) was measured at 450 nm. For IgG1 and IgG2c, the plates were developed with 1-Step^™ ABTS Substrate Solution (Thermo Scientific), and OD was measured at 405nm.