96-well plates (Corning) were coated with 2 ug/mL of recombinant SARS-CoV-2 RBD protein diluted in PBS and incubated at 4°C overnight. Plates were washed with PBS-T (PBS containing 0.05% Tween-20) and incubated with blocking buffer (PBS-T and 3% milk) for 1h at room temperature (RT). Plasma, culture supernatants or monoclonal antibodies were serially diluted in dilution buffer (PBS-T and 1% milk) in triplicate, added to plates, and incubated at RT for 2 h. Secondary antibodies were diluted in dilution buffer as follows: anti-human IgG-HRP (Jackson ImmunoResearch) at 1:3000 or anti-human IgA-HRP (Southern Biotech) at 1:1500. Plates were incubated with secondary antibodies for 1h at RT, then detected with 1X 3,30,5,50-Tetramethylbenzidine (TMB) (Invitrogen) and quenched with 1M HCl. Sample optical density (OD) was measured by a spectrophotometer at 450 nm and 570 nm.
Anti human iga hrp
Manufactured by Southern Biotech
Anti-human IgA-HRP is a conjugate of anti-human IgA antibody and horseradish peroxidase (HRP). It is used as a detection reagent in immunoassays to identify and measure the presence of human IgA antibodies in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Anti human igg hrp, by Jackson ImmunoResearch (3 mentions)
96 well plate, by Corning (3 mentions)
Anti human igm hrp, by Southern Biotech (2 mentions)
Human igg elisa kit, by STEMCELL (2 mentions)
Tetramethylbenzidine tmb, by Thermo Fisher Scientific (1 mentions)
Anti mouse igg1 hrp, by Southern Biotech (1 mentions)
Anti mouse igg2a hrp, by Southern Biotech (1 mentions)
Anti mouse igg hrp, by Southern Biotech (1 mentions)
Anti human igg hrp, by Southern Biotech (1 mentions)
Anti mouse iga hrp, by Southern Biotech (1 mentions)
3 3 5 5 tetramethylbenzidine (tmb), by Thermo Fisher Scientific (1 mentions)
Microplate spectrophotometer, by Agilent Technologies (1 mentions)
3 3 5 5 tetramethylbenzidine (tmb), by LGC (1 mentions)
Hydrochloric acid (hcl), by LGC (1 mentions)
5 protocols using anti human iga hrp
1
SARS-CoV-2 RBD Protein Binding Assay
2
SARS-CoV-2 Antibody Quantification in Mice and Humans
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A semi-quantitative ELISA was used to determine the levels of IgG, IgG1, IgG2a, and IgA antibodies in the sera of vaccinated mice. Also, the levels of IgG and IgA antibodies were determined in the sera of recovered COVID-19 patients.
First, high binding ELISA plates (Biomat, Italy) were coated with SARS-CoV-2 S- protein recombinant antigen (Sigma-Aldrich) at 1 μgmL−1. After washing plates, 50 μL of diluted serum samples collected from immunized mice and recovered COVID-19 patients were separately added to wells. After incubation for 1 h at 37 °C, plates were washed with PBS and then 100 μL of the following secondary antibody was separately added: 1) anti-mouse IgG-HRP, 2) anti-mouse IgG1-HRP, 3) anti-mouse IgG2a-HRP, 4) anti-mouse IgA-HRP, 5) anti-human IgG-HRP, 6) anti-human IgA-HRP (Southern Biotech). After incubation and washing with PBS, 50 μL of 3,3′, 5,5′-tetramethylbenzidine was added and then the reactions was stopped by adding 50 μL of 10% sulfuric acid. Finally, the absorbance of each well was read by a Spectrophotometer at 450 nm (BioTek Industries) and serum levels of antibodies was measured using a standard curve.
First, high binding ELISA plates (Biomat, Italy) were coated with SARS-CoV-2 S- protein recombinant antigen (Sigma-Aldrich) at 1 μgmL−1. After washing plates, 50 μL of diluted serum samples collected from immunized mice and recovered COVID-19 patients were separately added to wells. After incubation for 1 h at 37 °C, plates were washed with PBS and then 100 μL of the following secondary antibody was separately added: 1) anti-mouse IgG-HRP, 2) anti-mouse IgG1-HRP, 3) anti-mouse IgG2a-HRP, 4) anti-mouse IgA-HRP, 5) anti-human IgG-HRP, 6) anti-human IgA-HRP (Southern Biotech). After incubation and washing with PBS, 50 μL of 3,3′, 5,5′-tetramethylbenzidine was added and then the reactions was stopped by adding 50 μL of 10% sulfuric acid. Finally, the absorbance of each well was read by a Spectrophotometer at 450 nm (BioTek Industries) and serum levels of antibodies was measured using a standard curve.
3
SARS-CoV-2 Spike Protein Binding ELISA
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96-well plates (Corning) were coated with 2 ug/mL of recombinant SARS-CoV-2 RBD or trimeric spike protein diluted in PBS and incubated at 4°C overnight. Plates were washed with PBS-T (PBS containing 0.05% Tween-20) and incubated with blocking buffer (PBS-T and 3% milk) for 1 hour at room temperature (RT). Serum, culture supernatants or monoclonal antibodies were serially diluted in dilution buffer (PBS-T and 1% milk) in triplicate, added to plates, and incubated at RT for 2 hours. Secondary antibodies were diluted in dilution buffer as follows: anti-human IgG-HRP (Jackson ImmunoResearch) at 1:3000, anti-human IgM-HRP (Southern Biotech) at 1:3000, or anti-human IgA-HRP (Southern Biotech) at 1:1500. Plates were incubated with secondary antibodies for 1 hour at RT, then detected with 1X TMB (Invitrogen) and quenched with 1M HCl. Sample optical density (OD) was measured by a spectrophotometer at 450nm and 570nm. CR3022, a human SARS-CoV antibody previously determined to cross-react with SARS-CoV-2 was used as a positive control. IgG in culture supernatants was measured using a Human IgG ELISA Kit (Stemcell) according to the manufacturer’s instructions. Data was analysed in Prism (GraphPad).
4
SARS-CoV-2 Spike Protein ELISA
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96-well plates (Corning) were coated with 2 ug/mL of recombinant SARS-CoV-2 RBD or trimeric spike protein diluted in PBS and incubated at 4°C overnight. Plates were washed with PBS-T (PBS containing 0.05% Tween-20) and incubated with blocking buffer (PBS-T and 3% milk) for 1 h at room temperature (RT). Plasma, culture supernatants or monoclonal antibodies were serially diluted in dilution buffer (PBS-T and 1% milk) in triplicate, added to plates, and incubated at RT for 2 h. Secondary antibodies were diluted in dilution buffer as follows: anti-human IgG-HRP (Jackson ImmunoResearch) at 1:3000, anti-human IgM-HRP (Southern Biotech) at 1:3000, or anti-human IgA-HRP (Southern Biotech) at 1:1500. Plates were incubated with secondary antibodies for 1 h at RT, then detected with 1X 3,3′,5,5′-Tetramethylbenzidine (TMB) (Invitrogen) and quenched with 1M HCl. Sample optical density (OD) was measured by a spectrophotometer at 450nm and 570nm. CR3022, a human SARS-CoV antibody previously determined to cross-react with SARS-CoV-2 was used as a positive control. IgG in culture supernatants was measured using a Human IgG ELISA Kit (Stemcell) according to the manufacturer’s instructions. Data was analyzed in Prism (GraphPad).
5
Peanut-specific IgA ELISA in Saliva Samples
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