Goat anti human igg fc hrp

Binding to the SARS-CoV-2 spike protein were evaluated using His-tagged materials (S1, RBD, and S2) purchased from ACRObiosystems (San Jose, CA). These materials were used to coat Nunc 96-well Immulon 4HBX plates (100 μL) at a concentration of 1 μg/mL. The plates were incubated 12–24 hours at 2–4° C. The plates were then washed three times with Wash Buffer (WB; 1XPBS containing 0.05% Tween-20). The wells were then blocked using 200 μL of Blocking Buffer (BB; 1XPBS containing 3% BSA). The plates were incubated 1–4 hours at room temperature. The wells were then washed three times using WB. Positive control antibodies or test samples were diluted in Assay Dilution Buffer (ADB; 1XPBS, 0.05% Tween-20 and 1% BSA) and 100 μL was added to each well. The plates were incubated for 60±10 minutes at room temperature on a shaking platform. The plates were then washed three times using WB. The secondary goat anti-human IgG(Fc)-HRP (Abcam, Cambridge, MA) was diluted 1:25,000 in ADB and 100 μL was added to each well. The plates were incubated for 60±10 minutes at room temperature on a shaking platform. Finally, 100 μL of Stop Solution (0.2M H2SO4) was added to the plates and the plates were read at 450–630 nm using a Molecular Devices SpectraMax 384 Plus (Promega, Madison, WI). Each sample was tested in triplicate.