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).
384 well elisa plates
Manufactured by Corning
The 384-well ELISA plates are a laboratory equipment designed for enzyme-linked immunosorbent assay (ELISA) experiments. They provide a high-throughput platform for conducting multiple biochemical analyses simultaneously in a compact format. The plates feature 384 individual wells, allowing for the efficient processing of a large number of samples or test conditions in a single experiment.
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Lab products found in correlation
4 protocols using 384 well elisa plates
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).
3
ELISA for Evaluating HIV-1 Env Peptide Binding
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Enzyme-linked immunosorbent assays (ELISA) were performed as previously described8 (link). 384-well ELISA plates (Corning Life Sciences) were coated with TNC proteins (TNC-S, TNC-L, fn 1–8, fn 6–8, fn A-D, fbg, FNall, FNall+fbg) (Figure 1A ), human albumin, or bovine serum albumin at 100 μg/ml in 0.1M NaHCO3 and incubated overnight at 4°C. Blocking was performed using Super Block (1X PBS, 4% whey protein, 15% goat serum, 0.5% Tween 20) for 1 hour at RT. HIV-1 Env peptides (including MN.3 gp120, MN.3 gp70 V3, and the linear peptides MN.V3, MN.V3 15-mer, MN.V3 321/322, MN.V3 326/327, and MN.V3 321/322/326/327) were titrated using a two-fold serial dilution in Super Block starting at 500 μg/ml for the linear peptides, 1,280 μg/mL for MN.3 gp120, and 1,000 μg/ml for MN.3 gp70 V3. An HIV-1 gp120 specific monoclonal antibody (CH22 or 16H3) was used as a primary antibody while detection was done using goat anti-human or anti-mouse HRP labeled (1:5,000) as a secondary antibody. Signal was then detected with SureBlue Reserve 3,3’,5,5’-tetramethylbenzidine (TMB) substrate (VWR) and absorbance recorded at 450nm using a Spectramax Plus spectrophotometer (Molecular Devices).
4
ELISA for Evaluating HIV-1 Env Peptide Binding
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