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

blue ice or RT

Predicted reactivity based on immunogen sequence: SARS-CoV Spike proteins: (63%)

WB: 2-5 μ g/mL; IF: 20 μ g/mL; IHC: 0.5 μ g/mL.

Antibody validated: Western Blot in human samples; Immunohistochemistry and Immunofluorescence in human samples. SARS-CoV-2 (COVID-19) Spike RBD Antibody can be used for the detection of SARS-CoV-2 (COVID-19) Spike protein in ELISA and WB. 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 RBD antibody is affinity chromatography purified via peptide column.

IgG

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

SARS-CoV-2 (COVID-19) Spike RBD 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 RBD 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 RBD Antibody, 9087, 2 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 (9087) includes site 501N that was mutated in all three variants. 9087 has low binding affinity for all three variants as compared to WT.

Figure 3 Western Blot Validation with SARS-CoV-2 (COVID-19) Spike RBD Recombinant Protein
Loading: 30 ng per lane of SARS-CoV-2 (COVID-19) Spike RBD recombinant protein, 10-303. Antibodies: SARS-CoV-2 (COVID-19) Spike RBD, 9087, 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.

Figure 5 Immunofluorescence Validation of SARS-CoV-2 (COVID-19) Spike RBD in 293 Transfected Cells
Immunofluorescent analysis of 4% paraformaldehyde-fixed Spike transfected 293 cells labeling SARS-CoV-2 (COVID-19) Spike RBD with 9087 at 20 ug/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).

Figure 7 Western Blot Validation with SARS-CoV-2 (COVID-19) Spike RBD+SD1 Recombinant Protein
Loading: 30 ng per lane of SARS-CoV-2 (COVID-19) Spike RBD+SD1 recombinant protein, 10-304. Antibodies: SARS-CoV-2 (COVID-19) Spike RBD, 9087, 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.

Figure 9 Western Blot Validation with SARS-CoV-2 (COVID-19) Spike RBD+SD1+SD2 Recombinant Protein
Loading: 30 ng per lane of SARS-CoV-2 (COVID-19) Spike RBD+SD1+SD2 recombinant protein, 10-305. Antibodies: SARS-CoV-2 (COVID-19) Spike RBD, 9087, 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.

Figure 11 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 RBD, 9087, 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. Lane 1: 2.5 µg/mL and Lane 2: 5 µg/mL.