Optimal dilutions for each application to be determined by the researcher.
Figure 1 Detection of SARS-CoV-2 Variant Proteins with Spike S1 Antibodies by Direct ELISA Coating Antigen: SARS-CoV-2 full length spike proteins, including WT, UK variant (B.1.1.7), SA variant (B.1.135) and Brazil (P.1). Dilution: 1-3000 ng/mL. Incubate at 4 &730, C overnight.Detection Antibodies: SARS-CoV-2 Spike S1 Antibody, 10-351, 0.1 &956,g/mL, incubate at RT for 1 hr.Secondary Antibodies: Goat anti-rabbit HRP at 1:20,000, incubate at RT for 1 hr.Spike S1 antibody (10-351) cannot detect SA variant (B.1.135) and Brazil (P.1), both of which carry more mutations in RBD as compared to UK variant (B.1.1.7).
Figure 2 Immunohistochemistry Validation of SARS-CoV-2 (COVID-19) Spike in COVID-19 Patient Lung Immunohistochemical analysis of paraffin-embedded COVID-19 patient lung tissue using anti- SARS-CoV-2 (COVID-19) Spike antibody (10-351, 3 ug/mL). Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT, antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4&730,C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin. Strong signal of SARS-COV-2 Spike protein was observed in the macrophages of COVID-19 patient lung, but not in non-COVID-19 patient lung.
Figure 3 Sandwich ELISA for SARS-CoV-2 (COVID-19) Matched Pair Spike S1 AntibodiesAntibodies: SARS-CoV-2 (COVID-19) Spike Antibodies, 10-351 and 10-706. A sandwich ELISA was performed using SARS-CoV-2 Spike S1 antibody (10-351, 2ug/ml) as capture antibody, the Spike S1 recombinant protein as the binding protein (97-087), and the anti-SARS-CoV-2 Spike S1 antibody (10-706, 1ug/ml) as the detection antibody. Secondary: Mouse anti-human IgG HRP conjugate (PM6727) at 1:10000 dilution. Detection range is from 0.03 ng to 300 ng. EC50 = 1.58 ng
Figure 4 Sandwich ELISA for SARS-CoV-2 (COVID-19) Matched Pair Spike S1 AntibodiesAntibodies: SARS-CoV-2 (COVID-19) Spike Antibodies, 10-351 and 9083-biotin. A sandwich ELISA was performed using SARS-CoV-2 Spike S1 antibody (10-351, 2ug/ml) as capture antibody, the Spike S1 recombinant protein as the binding protein (97-087), and the anti-SARS-CoV-2 Spike S1 antibody (9083-biotin, 1ug/ml) as the detection antibody. Secondary: Streptavidin-HRP at 1:10000 dilution. Detection range is from 0.03 ng to 300 ng. EC50 = 1.96 ng
Figure 5 Sandwich ELISA for SARS-CoV-2 (COVID-19) Matched Pair Spike S1 AntibodiesAntibodies: SARS-CoV-2 (COVID-19) Spike Antibodies, 10-351 and 9091-biotin. A sandwich ELISA was performed using SARS-CoV-2 Spike S1 antibody (10-351, 2ug/ml) as capture antibody, the Spike S1 recombinant protein as the binding protein (97-087), and the anti-SARS-CoV-2 Spike S1 antibody (9091-biotin, 1ug/ml) as the detection antibody. Secondary: Streptavidin-HRP at 1:10000 dilution. Detection range is from 0.03 ng to 300 ng. EC50 = 3.14 ng
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