To test whether the RBD coupled on CuMVTT can be recognized by anti-RBD antibodies and ACE2, plates were first coated with 5 μg/mL anti-CuMVTT antibody (house-made monoclonal antibody from hybridoma) to capture the CuMVTT vaccine, followed by incubation with different concentrations of the CuMVTT–RBD vaccine. Then human anti-RBD antibody (Sanyou Biopharmaceuticals, Shanghai, China) or biotinylated ACE2 (Sino Biological, Beijing, China) was added and incubated on plates for 1 h at room temperature. Next, HRP-labelled goat anti-human IgG (Nordic-MUbio, Susteren, The Netherlands) or HRP-labelled streptavidin (Thermo Fisher Scientific) was added for 1 h at room temperature. The plates were developed as described above using a TMB substrate. The readout of OD450nm was plotted in accordance with increasing CuMVTT–RBD concentrations.
Hrp labelled streptavidin
Manufactured by Thermo Fisher Scientific
HRP-labelled streptavidin is a protein complex that consists of the biotin-binding protein streptavidin conjugated with the enzyme horseradish peroxidase (HRP). This complex can be used as a detection reagent in various immunoassay and other biotechnological applications.
Automatically generated - may contain errors
Lab products found in correlation
Biotinylated ace2, by Sino Biological (1 mentions)
Anti p65 antibody, by Cell Signaling Technology (1 mentions)
Poly lactic co glycolic acid, by Merck Group (1 mentions)
Tgf β elisa kit, by R&D Systems (1 mentions)
Anti a20 antibody, by Abcam (1 mentions)
Anti β actin antibody, by Cell Signaling Technology (1 mentions)
Cholera toxin, by Merck Group (1 mentions)
Tnf α, by R&D Systems (1 mentions)
Ifn γ, by R&D Systems (1 mentions)
2 protocols using hrp labelled streptavidin
1
CuMV-RBD Vaccine Binding Assay
2
Murine Cytokine and Immune Cell Analysis
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Biotin-labelled sheep anti-mouse IgE antibody (cat. no. 1110-08) was purchased from Southern Biotech. HRP-labelled sheep anti-mouse IgG1 antibody (cat. no. A10551) and HRP-labelled sheep anti-mouse IgG2a antibody (cat. no. A10685) was purchased from Life Technologies; Thermo Fisher Scientific, Inc. HRP-labelled streptavidin (cat. no. A0303) was obtained from Beyotime Institute of Biotechnology. Mouse IL-4, IL-10, IL-13, TNF-α, IFN-γ and TGF-β ELISA Kits were purchased from R&D Systems. Cholera toxin and poly (lactic-co-glycolic acid) were purchased from SigmaAldrich; Merck KGaA. APC rat anti-mouse CD4 antibody (cat. no. 553051), FITC rat anti-mouse CD25 antibody (cat. no. 558689) and PE rat anti-mouse Foxp3 (cat. no. 560408) were obtained from BD Pharmingen; BD Biosciences. Anti-A20 antibody (product code ab74037) was purchased from Abcam, and anti-p65 antibody (product no. 3033S) and anti-β-actin antibody (4967S) were obtained from Cell Signaling Technology. The 293T cell line (CBP60439) was obtained from Cobioer.
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