Three hundred eighty-four-well ELISA plates (Maxisorp) were coated with 1 μg/ml purified GST-Jo-1 or GST protein in 1X PBS overnight at 4 °C. Plates were blocked with 5% FBS in 1X PBS plus 0.5% Tween (1X PBS-T) at 25 °C and washed 5X with 1X PBS-T. Plates were incubated at RT with 1:1000 patient sera or stimulated PBMC culture supernatants diluted 1:2 in blocking buffer. Parallel wells were incubated with 100X soluble GST-Jo-1 competitor during sera binding to measure specific binding. Bound antibody was detected by anti-human IgG Fc-specific-HRP (Sigma), anti-human IgG-HRP, or anti-human IgM-HRP (Southern Biotech) secondary antibodies diluted in 5% FBS/1X PBS-T. Plates were washed, TMB Ultra ELISA substrate (Thermo Fisher) was added, and plates were read at O.D. 370 nm using a microplate reader (SpectraMax M3).
Anti human igm hrp
Manufactured by Southern Biotech
Anti-human IgM-HRP is a laboratory reagent that consists of anti-human IgM antibodies conjugated with horseradish peroxidase (HRP). It is used as a detection tool in various immunoassay techniques, such as ELISA, to measure and quantify the presence of human IgM antibodies in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Anti human igg hrp, by Jackson ImmunoResearch (2 mentions)
96 well plate, by Corning (2 mentions)
Anti human iga hrp, by Southern Biotech (2 mentions)
Human igg elisa kit, by STEMCELL (2 mentions)
Ultra tmb elisa substrate, by Thermo Fisher Scientific (1 mentions)
Anti human igg fc specific hrp, by Merck Group (1 mentions)
Anti human igg hrp, by Southern Biotech (1 mentions)
3 3 5 5 tetramethylbenzidine (tmb), by Thermo Fisher Scientific (1 mentions)
3 protocols using anti human igm hrp
1
GST-Jo-1 ELISA for Autoantibody Detection
2
SARS-CoV-2 Spike Protein Binding ELISA
Check if the same lab product or an alternative is used in the 5 most similar protocols
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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).
3
SARS-CoV-2 Spike Protein ELISA
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
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).
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