The antibody tilters in sera from pre, first and second bleeds were determined using direct coating ELISA. The 384-well ELISA plates (Corning) were coated with 3 μg/mL SARS-CoV-2 RBD-his tag protein (Sino) in PBS at 4°C overnight. After standard washing with PBS-T washing buffer (phosphate-buffered saline containing 0.05% Tween 20), ELSIA plates were blocked with blocking buffer (2% bovine serum albumin dissolved in PBST and filtered) for 1 hr at room temperature. Serial dilutions of pre-immune, first and second immune anti-sera in blocking buffer were added into plates and for 1hr at room temperature. Plates were washed and incubated with relative goat anti-mouse IgG(H+L)/HRP (Thermo Fisher,1:5000) for 1 h at room temperature. Plates were washed and developed using TMB reagents as substrates (Biolegend) following the manufacturer’s recommended protocol. Reaction was stop with stop solution (1M H3PO4) and absorbance at 450nm was recorded by a microplate reader (Perkin Elmer).
Relative goat anti mouse igg h l hrp
Manufactured by Thermo Fisher Scientific
Relative goat anti-mouse IgG(H+L)/HRP is a secondary antibody conjugated with horseradish peroxidase (HRP). It is designed to detect and bind to mouse immunoglobulins (IgG) in a variety of immunoassays and immunochemical procedures.
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Lab products found in correlation
2 protocols using relative goat anti mouse igg h l hrp
1
SARS-CoV-2 RBD Antibody Titer Quantification
2
SARS-CoV-2 RBD Antibody Titer Analysis
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The antibody tilters in sera from pre, first, and second bleeds were determined using direct coating ELISA. The 384-well ELISA plates (Corning) were coated with 3 μg/mL SARS-CoV-2 RBD-his tag protein (Sino) in PBS at 4 °C overnight. After standard washing with PBST washing buffer (phosphate-buffered saline containing 0.05% Tween 20), ELISA plates were blocked with blocking buffer (2% bovine serum albumin dissolved in PBST and filtered) for 1 h at room temperature. Serial dilutions of pre-immune, first, and second immune anti-sera in blocking buffer were added into plates and for 1 h at room temperature. Plates were washed and incubated with relative goat anti-mouse IgG(H + L)/HRP (Thermo Fisher,1:5000) for 1 h at room temperature. Plates were washed and developed using TMB reagents as substrates (Biolegend) following the manufacturer’s recommended protocol. The reaction was stopped with a stop solution (1 M H3PO4) and absorbance at 450 nm was recorded by a microplate reader (Perkin Elmer).
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