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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: 10-3000 ng/mL. Incubate at 4 ˚ C overnight.
Detection Antibodies: SARS-CoV-2 Spike S1 Antibody, 9083, 1 μg/mL, incubate at RT for 1 hr.
Secondary Antibodies: Goat anti-rabbit HRP at 1:20,000, incubate at RT for 1 hr.
Immunogen region of antibody (9083) includes sites 20T and 26P that were mutated in Brazil variant P.1.. Therefore, Spike S1 Antibody (9083) cannot detect P.1 variant. See high quality image.
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Figure 2 Immunohistochemistry Validation of SARS-CoV-2 (COVID-19) Spike S1 in Human Lung Tissue from the COVID-19 Patient (Sun et al., 2020)
Detection of SARS-CoV-2 nucleocapsid protein by anti-SARS-COV-2 nucleocapsid antibodies (9099, 0.02 μg/mL, A,B) or SARS-CoV-2 Spike S1 antibodies (9083, 1 μg/mL D,E) in adjacent sections of autopsy lung tissue from COVID-19 deceased patient. Negative control staining on autopsy lung tissue from patient who died from non-COVID-19 pneumonia is shown for Nucleocapsid protein (C) or Spike protein (F). Negative control using normal rabbit immunoglobulin on COVID-19 autopsy tissue is presented (G). DAB chromogen and hematoxylin counterstain are used. Scale bars: 50μM in A, C, D, F, G; 20μM in B and E. See high quality image.
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Figure 3 Immunohistochemistry Validation of SARS-CoV-2 (COVID-19) Spike S1 in COVID-19 Patient Lung
Immunohistochemical analysis of paraffin-embedded COVID-19 patient lung tissue using anti- SARS-CoV-2 (COVID-19) Spike S1 antibody (9083, 0.5 μg/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 ˚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 macrophage of COVID-19 patient lung, but not in non-COVID-19 patient lung. See high quality image.
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Figure 4 Immunohistochemistry Validation of SARS-CoV-2 (COVID-19) Spike S1 in COVID-19 Patient Lung
Immunohistochemical analysis of paraffin-embedded COVID-19 patient lung tissue using anti- SARS-CoV-2 (COVID-19) Spike S1 antibody (9083, 0.5 μg/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 ˚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 macrophage of COVID-19 patient lung. See high quality image.
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Figure 5 Immunohistochemistry Validation of SARS-CoV-2 (COVID-19) Spike S1 in Human Lung Tissue from the COVID-19 Patient
Immunohistochemical analysis of paraffin-embedded COVID-19 patient lung tissue using anti- SARS-CoV-2 (COVID-19) Spike S1 antibody (9083, 1 μg/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˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin. (Courtesy of Hallgeir Rui, MCW) (Picture shown in 40X magnification) See high quality image.
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Figure 6 Immunohistochemistry Validation of SARS-CoV-2 (COVID-19) Spike S1 in Human Lung Tissue from the COVID-19 Patient
Immunohistochemical analysis of paraffin-embedded COVID-19 patient lung tissue using anti- SARS-CoV-2 (COVID-19) Spike S1 antibody (9083, 1 μg/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˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin. (Courtesy of Hallgeir Rui, MCW) (Picture shown in 40X magnification) See high quality image.
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Figure 7 Immunohistochemistry Validation of SARS-CoV-2 (COVID-19) Spike S1 in Human Lung Tissue from the COVID-19 Patient
Immunohistochemical analysis of paraffin-embedded COVID-19 patient lung tissue using anti- SARS-CoV-2 (COVID-19) Spike S1 antibody (9083, 1 μg/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˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin. (Courtesy of Hallgeir Rui, MCW) (Picture shown in 40X magnification) See high quality image.
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Figure 8 Immunohistochemistry Validation of SARS-CoV-2 (COVID-19) Spike S1 in Human Lung Tissue from the COVID-19 Patient
Immunohistochemical analysis of paraffin-embedded COVID-19 patient lung tissue using anti- SARS-CoV-2 (COVID-19) Spike S1 antibody (9083, 1 μg/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˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin. (Courtesy of Hallgeir Rui, MCW) (Picture shown in 40X magnification) See high quality image.
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Figure 9 Immunohistochemistry Validation of SARS-CoV-2 (COVID-19) Spike S1 in Human Lung Tissue from the COVID-19 Patient
Immunohistochemical analysis of paraffin-embedded COVID-19 patient lung tissue using anti- SARS-CoV-2 (COVID-19) Spike S1 antibody (9083, 1 μg/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˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin. (Courtesy of Hallgeir Rui, MCW) (Picture shown in 20X magnification) See high quality image.
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Figure 10 Immunohistochemistry Validation of SARS-CoV-2 (COVID-19) Spike S1 in Human Lung Tissue from the COVID-19 Patient
Immunohistochemical analysis of paraffin-embedded COVID-19 patient lung tissue using anti- SARS-CoV-2 (COVID-19) Spike S1 antibody (9083, 1 μg/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˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin. (Courtesy of Hallgeir Rui, MCW) (Picture shown in 20X magnification) See high quality image.
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Figure 11 Overexpression Validation in Spike Transfected 293 Cells
Loading: 10 μg per lane of 293 cell lysate. Antibodies: SARS-CoV-2 (COVID-19) Spike S1, 9083 (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. Lane 1: WT 293 cells and Lane 2: SARS-CoV-2 Spike overexpressed 293 cells See high quality image.
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Figure 12 Immunofluorescence Validation of SARS-CoV-2 (COVID-19) Spike S1 in 293 Transfected Cells
Immunofluorescent analysis of 4% paraformaldehyde-fixed Spike transfected 293 cells labeling SARS-CoV-2 (COVID-19) Spike S1 with 9083 at 5 ug/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue). See high quality image.
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Figure 13 ELISA Validation with SARS-CoV-2 (COVID-19) Spike Recombinant Protein
Antibodies: SARS-CoV-2 (COVID-19) Spike S1 antibody, 9083. A direct ELISA was performed using SARS-CoV-2 (COVID-19) Spike S1 recombinant protein (97-087) as coating antigen and the anti-SARS-CoV-2 (COVID-19) Spike S1 antibody as the capture antibody. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:20000 dilution. Detection range is from 8 ng/mL to 1000 ng/mL. See high quality image.
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Figure 14 Western Blot Validation with SARS-CoV-2 (COVID-19) Spike Recombinant Protein
Loading: 50 ng per lane of SARS-CoV-2 (COVID-19) Spike S1 recombinant protein (97-087). Antibodies: SARS-CoV-2 (COVID-19) Spike 1, 9083, 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. Lane 1: 1 μg/mL and Lane 2: 2 μg/mL See high quality image.
Specifications
