Hrp conjugated goat anti mouse igg h l antibody

HRP-conjugated goat anti-mouse IgG (H + L) antibody and HRP-conjugated goat anti-rabbit IgG (H + L) antibody (Jackson ImmunoResearch Laboratories, United States).

Edmonston strain-derived MV antigens (Jena Bioscience) or recombinant S protein (ABIN6952426, Antibodies Online) were coated on NUNC MAXISORP 96-well immuno-plates (Thermo Fisher) at 1 µg/ml. Coated plates were incubated overnight at 4 °C. After washing and blocking, sera from immunized mice were serially diluted in the binding buffer and incubated on plates for 1 h at 37 °C. After washing steps, an HRP-conjugated goat anti-mouse IgG (H + L) antibody (Jackson ImmunoResearch, 115-035-146, 1:5000 dilution) was added for 1 h at 37 °C. Antibody binding was detected by the addition of the TMB substrate (Eurobio) and the reaction was stopped with 100 µl of 30% H₂SO₄. The optical densities were recorded at 450 and 620 nm wavelengths using the EnSpire 2300 Multilabel Plate Reader (Perkin Elmer). Endpoint titers for each individual serum sample were calculated as the reciprocal of the last dilution giving twice the absorbance of the negative control sera. Isotype determination of the antibody responses was performed using HRP-conjugated isotype-specific (IgG1 or IgG2a) goat anti-mouse antibodies (AB97240 and AB97245, Abcam, 1:5000).