20μl of ectodomains (stabilized prefusion trimer) of SARS-CoV-2 or SARS-CoV, or the disulphide stabilized SARS-CoV-2 or SARS-CoV, were coated on 384 well ELISA plates at 1 ng/μl for 16 hours at 4°C. Plates were washed with a 405 TS Microplate Washer (BioTek Instruments) then blocked with 80 μl SuperBlock (PBS) Blocking Buffer (Thermo Scientific) for 1 hour at 37°C. Plates were then washed and 30 μl antibodies were added to the plates at concentrations between 4 × 10−8 and 10 ng/μl and incubated for 1 h at 37°C. Plates were washed and then incubated with 30 μl of 1/5000 diluted goat anti-human Fc IgG-HRP (invitrogen A18817). Plates were washed and then 30 μl Substrate TMB microwell peroxidase (Seracare 5120–0083) was added for 5 min at room temperature. The colorimetric reaction was stopped by addition of 30 μl of 1 N HCl. A450 was read on a Varioskan Lux plate reader (Thermo Scientific).
Goat anti human fc igg hrp
Manufactured by Thermo Fisher Scientific
Goat anti-human Fc IgG-HRP is a secondary antibody conjugated with horseradish peroxidase (HRP). It is designed to detect and quantify human immunoglobulin G (IgG) in various immunoassay applications.
Automatically generated - may contain errors
2 protocols using goat anti human fc igg hrp
1
SARS-CoV-2 Spike Protein ELISA Assay
2
Amyloid-beta Fibril Characterization
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Fibrils were also assembled using Aβ peptides with N-terminal deletions including Aβ2-42 (Bachem, 40306028.0500), Aβ3-42 (Bachem, 4090137.0500), Aβ4-42 (Bachem, 4090138.0500), Aβ5-42 (Bachem, 4041241.0500), and Aβ11-42 (Anaspec, 63317) in addition to Aβ1-42 and purified using ultracentrifugation. Fibrils were then spotted on nitrocellulose membranes at equal Thioflavin T florescence. Membranes were blocked with 5% milk in PBS at room temperature for 1 h followed by 3x washing with PBST (PBS with 0.1% Tween 20). Membranes were then incubated with Aβ antibodies at 10 nM (1% milk) in PBST at room temperature for 2 to 3 h. Following primary incubation, membranes were washed 3x with PBST followed by incubation with goat anti-human Fc IgG HRP (1/5000x dilution, Invitrogen, A18817) in PBST at room temperature (1 h). Following secondary incubation, the blots were washed 3x with PBST, developed with ECL (Pierce, 32109), and imaged with a BioRad imager.
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