30 μL of 3 ug/mL of SARS-CoV-2 S, NTD, RBD, S1 S2(Pre), or S2(Post) diluted in PBS were incubated on a 384-well Nunc Maxisorp plate (ThermoFisher 464718) for one hour at 37°C. Plates were slapped dry before addition of 80 μL blocker Casein in PBS (ThermoFisher) and incubation for one hour at 37°C. Plates were slapped dry and a 1:4 serial dilution of plasma in 30 μL TBST was added and incubated for one hour at 37°C. Plates were slapped dry and washed 4x with TBST using a BioTek plate washer followed by addition of Invitrogen anti-Human IgG (ThermoFisher A18817) and one hour incubation at 37°C. Plates were once again slapped dry and washed 4x with TBST before addition of room temperature TMB Microwell Peroxidase (Seracare 5120–0083). The reaction was quenched after 1–2 minutes with 1 N HCl and the A450 of each well was read using a BioTek plate reader. Prism (GraphPad) area under curve (AUC) was used to analyze data following log transformation of dilution series.
Invitrogen anti human igg
Manufactured by Thermo Fisher Scientific
The Invitrogen anti-Human IgG is a laboratory reagent used to detect and quantify human immunoglobulin G (IgG) in samples. It is designed for use in various immunoassay techniques.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using invitrogen anti human igg
1
SARS-CoV-2 Spike Protein ELISA
2
SARS-CoV-2 Spike Protein Binding Assay
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30 μL of 3 ug/mL of SARS-CoV-2 S Hexapro, NTD, RBD, S2(Pre), or S2(Post) diluted in PBS were incubated on a 384-well Nunc Maxisorp plate (ThermoFisher 464718) for one hour at 37C. Plates were slapped dry before addition of 80 μL blocker Casein in PBS (ThermoFisher) and incubation for one hour at 37C. Plates were slapped dry and a 1:4 serial dilution of plasma in 30 μL TBST was added and incubated for one hour at 37C. Plates were slapped dry and washed 4x with TBST using a BioTek plate washer followed by addition of Invitrogen anti-Human IgG (ThermoFisher A18817) and one hour incubation at 37C. Plates were once again slapped dry and washed 4x with TBST before addition of room temperature TMB Microwell Peroxidase (Seracare 5120–0083). The reaction was quenched after 1–2 minutes with 1 N HCl and the A450 of each well was read using a BioTek plate reader. Prism (GraphPad) nonlinear regression with “Sigmoidal, 4PL, X is concentration” was used to determine the ED50 of each sample. The bottom was constrained to the minimum A450 per plate and top was constrained to the maximum A450 per plate, besides postfusion S binding for Ad26.COV2.S and Sputnik V where the top was constrained to less than the maximum A450 per plate due to wide variability in maximum A450.
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