SARS-CoV-2 (COVID-19) Spike S1 Antibody

blue ice or RT

Predicted reactivity based on immunogen sequence: SARS-CoV Spike proteins: (44%)

IHC: 0.5 μ g/mL; WB: 1 μ g/mL; IF: 5 μ g/mL;

Antibody validated: Western Blot in human samples; Immunofluorescence in human samples; Immunohistochemistry in human samples. SARS-CoV-2 (COVID-19) Spike S1 antibody can be used for the detection of SARS-CoV-2 (COVID-19) Spike protein in ELISA. It will detect 4 ng of free peptide at 1 μ g/mL. All other applications and species not yet tested.

SARS-CoV-2 (COVID-19) Spike S1 antibody is affinity chromatography purified via peptide column.

IgG

SARS-CoV-2 (COVID-19) Spike S1 antibody is supplied in PBS containing 0.02% sodium azide.

SARS-CoV-2 (COVID-19) Spike S1 antibody can be stored at 4˚ C for three months and -20˚ C, stable for up to one year. As with all antibodies care should be taken to avoid repeated freeze thaw cycles. Antibodies should not be exposed to prolonged high temperatures.

Optimal dilutions/concentrations should be determined by the end user. The information provided is a guideline for product use. This product is for research use only.

S

1791269090

Optimal dilutions for each application to be determined by the researcher.

surface glycoprotein

Coronavirus disease 2019 (COVID-19), formerly known as 2019-nCoV acute respiratory disease, is an infectious disease caused by SARS-CoV-2, a virus closely related to the SARS virus (1). The disease is the cause of the 2019–20 coronavirus outbreak (2). The structure of 2019-nCoV consists of the following: a Spike protein (S), hemagglutinin-esterease dimer (HE), a membrane glycoprotein (M), an envelope protein (E) a nucleoclapid protein (N) and RNA. Coronavirus invades cells through Spike (S) glycoproteins, a class I fusion protein. It is the major viral surface protein that coronavirus uses to bind to the human cell surface receptor. It also mediates the fusion of host and viral cell membrane, allowing the virus to enter human cells and begin infection (3). The spike protein is the major target for neutralizing antibodies and vaccine development (4). The protein modeling suggests that there is strong interaction between Spike protein receptor-binding domain and its host receptor angiotensin-converting enzyme 2 (ACE2), which regulate both the cross-species and human-to-human transmissions of COVID-19 (5). The recent study has shown that the SARS-CoV-2 spike protein binds ACE2 with higher affinity than SARS-CoV spike protein (6).

Hui et al. Int J Infect Dis. 2020; 91:264-266.

Lee et al. J Virol. 2006; 80(8):4079-87.

Wrapp et al. Science. 2020.

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 μ g/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.

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 μ g/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.

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 μ g/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)

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 μ g/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)

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 μ g/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)

Figure 11 Overexpression Validation in Spike Transfected 293 Cells
Loading: 10 μ g per lane of 293 cell lysate. Antibodies: SARS-CoV-2 (COVID-19) Spike S1, 9083 (1 μ g/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

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.