6d8 directed against ecd3

The ELISA was performed using a rabbit polyclonal anti-human desmoglein-2 antibody (R&D System, Catalog # AF947) as a capture antibody at a concentration of 2 µg/ml in 0.1 M Na-carbonate, pH = 9.6 buffer. Plates were washed with TBS-T (Tris-buffered saline + 0.05% Tween-20) before blocking with Starting Block^TM (PBS) Blocking Buffer (Thermo Scientific, Prod #37538). Human serum samples were analyzed in three dilutions (1:10, 1:50, and 1:100) using Blocking Buffer as a dilutant. For detection, the mouse monoclonal antibody 6D8 directed against ECD3 (AbD Serotec) was used at a 1:100 dilution. After three washes, goat anti-mouse IgG-HRP peroxidase (BD Pharmingen) was added at a 1:1000 dilution in blocking buffer. 1-Step^TM Ultra TMB-ELISA (Thermo Scientific) was used as a substrate. Recombinant human-DSG2 protein used for the standard curve was from Leinco Technologies, Inc. The detection limit of the DSG2 ELISA was 0.5 ng/ml.

ELISA was performed using the goat polyclonal anti-DSG2 antibody AF947 (R&D Systems, Minneapolis, MN) and the mouse monoclonal antibody 6D8 directed against ECD3 (AbD Serotec, Raleigh, NC). The detection limit of the DSG2 ELISA was 0.5 ng/ml. The antibodies cross-react with macaque DSG2.