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

Product Code: PSI-9087

Supplier: ProSci

Code	Size	Price

PSI-9087-0.02mg

0.02mg

£150.00

Quantity:

PSI-9087-0.1mg

0.1mg

£449.00

Quantity:

Prices exclude any Taxes / VAT

Overview

Host Type: Rabbit

Antibody Isotype: IgG

Antibody Clonality: Polyclonal

Regulatory Status: RUO

Target Species: Virus

Applications:

Enzyme-Linked Immunosorbent Assay (ELISA)
Immunofluorescence (IF)
Western Blot (WB)

Storage:

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.

Images

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<strong> Figure 1 Detection of SARS-CoV-2 Variant Proteins with Spike RBD Antibodies by Direct ELISA</strong><br>
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 μg/mL, incubate at RT for 1 hr.
Secondary Antibodies: Goat anti-rabbit HRP at 1:20,000, incubate at RT for 1 hr.
<strong>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. </strong>

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<strong>Figure 2 Immunohistochemistry Validation of SARS-CoV-2 (COVID-19) Spike RBD in COVID-19 Patient Lung</strong><br>
Immunohistochemical analysis of paraffin-embedded COVID-19 patient lung tissue using anti- SARS-CoV-2 (COVID-19) Spike RBD antibody (9087, 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 RBD protein was observed in macrophage of COVID-19 patient lung, but not in non-COVID-19 patient lung.

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<strong>Figure 3 Western Blot Validation with SARS-CoV-2 (COVID-19) Spike RBD Recombinant Protein </strong><br>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 ug/mL and Lane 2: 2 ug/mL.

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<strong>Figure 4 ELISA Validation with SARS-CoV-2 (COVID-19) Spike RBD Recombinant Protein</strong><br>Antibodies: SARS-CoV-2 (COVID-19) Spike RBD antibody, 9087 (1 μg/mL). A direct ELISA was performed using SARS-CoV-2 (COVID-19) Spike RBD recombinant protein (10-303) as coating antigen and the anti-SARS-CoV-2 (COVID-19) Spike RBD antibody as the capture

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<strong>Figure 5 Immunofluorescence Validation of SARS-CoV-2 (COVID-19) Spike RBD in 293 Transfected Cells</strong><br>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).

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<strong>Figure 6 Overexpression Validation in Spike Transfected 293 Cells </strong><br>Loading: 10 μg per lane of 293 cell lysate. Antibodies: SARS-CoV-2 (COVID-19) Spike RBD, 9087 (4 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

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<strong>Figure 7 Western Blot Validation with SARS-CoV-2 (COVID-19) Spike RBD+SD1 Recombinant Protein</strong><br>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 ug/mL and Lane 2: 2 ug/mL.

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<strong>Figure 8 ELISA Validation with SARS-CoV-2 (COVID-19) Spike RBD+SD1 Recombinant Protein</strong><br>Antibodies: SARS-CoV-2 (COVID-19) Spike RBD antibody, 9087 (1 μg/mL). A direct ELISA was performed using SARS-CoV-2 (COVID-19) Spike RBD+SD1 recombinant protein (10-304) as coating antigen and the anti-SARS-CoV-2 (COVID-19) Spike RBD antibody as the capture antibody. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:20000 dilution. Detection range is from 8 ng/mL to 1000ng/mL.

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<strong>Figure 9 Western Blot Validation with SARS-CoV-2 (COVID-19) Spike RBD+SD1+SD2 Recombinant Protein</strong><br>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 ug/mL and Lane 2: 2 ug/mL.

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<strong>Figure 10 ELISA Validation with SARS-CoV-2 (COVID-19) Spike Recombinant Protein</strong><br>Antibodies: SARS-CoV-2 (COVID-19) Spike antibody, 9087 (1 μg/mL). 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 RBD antibody as the capture antibody. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:20000 dilution. Detection range is from 16 ng/mL to 2000ng/mL.

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<strong>Figure 11 Western Blot Validation with SARS-CoV-2 (COVID-19) Spike Recombinant Protein </strong><br>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 ug/mL and Lane 2: 5 ug/mL.

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Documents

Further Information

Additional Names:

SARS-CoV-2 (COVID-19) Spike RBD Antibody: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), Surface Glycoprotein, Spike protein

Application Note:

WB: 2-5 μg/mL; IF: 20 μg/mL; IHC: 0.5 μg/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 μg/mL. All other applications and species not yet tested.

Background:

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).

Background References:

Gorbalenya. bioRxiv: 2020.
Hui et al. Int J Infect Dis. 2020;91:264-266.
Belouzard et al. Viruses. 2012;4(6):1011-33.
Lee et al. J Virol. 2006;80(8):4079-87.
Wan et al. J Virol. 2020.
Wrapp et al. Science. 2020.

Buffer:

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

Concentration:

1 mg/mL

Conjugate:

Unconjugated

DISCLAIMER:

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.

Homology:

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

Immunogen:

Anti-SARS-CoV-2 (COVID-19) Spike RBD antibody (9087) was raised against a peptide corresponding to 19 amino acids near the carboxy?terminus of SARS-CoV-2 (COVID-19) Spike glycoprotein RBD.

The immunogen is located within the last 50 amino acids of SARS-CoV-2 (COVID-19) Spike protein RBD.

ISOFORMS:

SARS-CoV-2 (COVID-19) Spike has one isoform (1273aa).

NCBI Gene ID #:

43740568

NCBI Official Name:

surface glycoprotein

NCBI Official Symbol:

NCBI Organism:

SARS-CoV-2

Physical State:

Liquid

Protein Accession #:

QHD43416

Protein GI Number:

1791269090

Purification:

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

Research Area:

Infectious Disease,COVID-19

Swissprot #:

P0DTC2

User NOte:

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

References

Heaton, et al. SRSF protein kinases 1 and 2 are essential host factors for human coronaviruses including SARS-CoV-2. bioRxiv. 2020 Aug 14;2020.08.14.251207. doi: 10.1101/2020.08.14.251207.PMID: 32817937