SARS Spike Antibody

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ProSci
Product Code: PSI-3525
Product Group: Primary Antibodies
Supplier: ProSci

CodeSizePrice
PSI-3525-0.02mg0.02mg£150.00
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PSI-3525-0.1mg0.1mg£449.00
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Overview

Host Type: Rabbit
Antibody Isotype: IgG
Antibody Clonality: Polyclonal
Regulatory Status: RUO
Application: Enzyme-Linked Immunosorbent Assay (ELISA)

Images

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<strong>Figure 1 Immunofluorescent Validation of 3525 in SARS-CoV-2 Infected Lung Tissue (Singh et al., Nature Microbiology, 2021) </strong><br>Multilabel confocal immunofluorescence microscopy of formalin-fixed paraffin-embedded lung sections from rhesus macaques infected with SARS-CoV-2. SARS-CoV-2 spike-specific antibodies, 3525 ProSci Inc. (k, n) (turquoise); Ki67 (magenta) and neutrophil marker CD66abce (yellow) (k); pan-macrophage marker CD68 (magenta) (n) and DAPI (blue).
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<strong> Figure 2 Immunofluorescent Validation of 3525 in SARS-CoV-2 Infected Nose and Tonsil (Singh et al., Nature Microbiology, 2021) </strong><br> Multi-label confocal immunofluorescence microscopy of nasal epithelium (20X-b, 63xh) and tonsil (20X-c,63X-i) from rhesus macaques infected with SARS-CoV-2 with SARS-CoV-2 spike-specific antibodies, 3525 ProSci Inc. (turquoise), DAPI (blue). Rabbit IgG isotype control antibody was used to stain the tissues to rule out any non-specific staining (e, f).
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<strong> Figure 3 Immunofluorescent Validation of 3525 in SARS-CoV-2 Infected Lung Tissue (Singh et al., Nature Microbiology, 2021) </strong><br> Multilabel confocal immunofluorescence microscopy of formalin-fixed paraffin-embedded lung sections from rhesus macaques infected with SARS-CoV-2. SARS-CoV-2 spike-specific antibodies, 3525 ProSci Inc. (k-s) (turquoise); Ki67 (magenta) and neutrophil marker CD66abce (yellow) (k-m); pan-macrophage marker CD68 (magenta) (n-p); HLA-DR (magenta) and pDC marker CD123 (yellow) (q?s) and DAPI (blue).
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<strong>Figure 4 Immunohistochemistry Validation of SARS-CoV-2 (COVID-19) Spike 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 S2 antibody (3525, 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 spike protein signal was observed in macrophages and airway epithelium of COVID-19 patient lung, but not in non-COVID-19 patient lung.
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<strong>Figure 5 Immunohistochemistry Validation of SARS-CoV-2 (COVID-19) Spike 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 S2 antibody (3525, 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 spike protein signal was observed in macrophages of COVID-19 patient lung.
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<strong>Figure 6 IHC/IF Validation in COVID-19 Patient Sample </strong><br> (Nuovo et al., 2020) Detection of SARS-CoV-2 proteins in nasopharyngeal swab cell preparations. B. An intense signal for covid-19 spike protein <strong> tested by SARS-CoV-2 spike antibodies (3525) </strong> was observed in the glandular cells. F-H. Co-expression of spike detected by spike antibodies (3525, 0.2 μg/mL) and envelope proteins detected by envelope antibodies (3531, 2 μg/mL) of SARS-CoV-2 (F) documented localization of each protein to glandular cells (G, yellow). No signal was seen in oral swabs of positive cases (H). <strong>Both the spike and envelope protein detected by anti-spike antibodies (3525) and anti-envelope antibodies (3531) produced a signal in the nasopharyngeal swabs of the three cases and no signal was evident in the nasopharyngeal swabs of the seven controls. </strong>
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<strong>Figure 7 IF Validation of SARS-CoV2 Spike in COVID-19 Patient Lung </strong><br> (Magro et al., 2020) SARS-CoV2 spike protein (red, panel C) detected detected by anti-spike antibodies (3525, 0.2 μg/mL) colocalized with C4d (green in panel d, merged in yellow). Spike protein (red, panel g) was also colocalized with C5b-9 (green in panel f&h, merged in yellow)
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<strong>Figure 8 IF Validation of SARS-CoV2 Spike in COVID-19 Patient Skin</strong><br> (Magro et al., 2020) C4d is highlighted green while COVID-19 spike protein detected by anti-spike antibodies (3525, 0.2 μg/mL) shows a red staining pattern; a yellow signal is discernible indicative of co-localization of C4d and viral protein within the microvasculature.
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<strong>Figure 9 IHC Validation of SARS-CoV2 Spike in the Nasopharyngeal Swab Sample of the COVID-19 Patient </strong><br> Strong spike signal was detected by anti-spike antibodies (3525, 0.2 μg/mL) in the nasopharyngeal swab sample of the COVID-19 patient and no spike signal was observed in the sample of the COVID-19 negative patient. COVID-19 cases were confirmed by PCR. (Courtesy of Dr. Nuovo Gerard J., OSU)
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<strong>Figure 10 IHC Validation of SARS-CoV2 Spike in COVID-19 Patient Lung</strong><br> Strong spike signal was detected by anti-spike antibodies (3525, 0.2 μg/mL) in the lung of the COVID-19 patient confirmed by PCR. (Courtesy of Dr. Nuovo Gerard J., OSU)
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<strong>Figure 11 IHC Validation of SARS-CoV2 Spike in COVID-19 Patient Brain</strong><br> Spike protein was detected by anti-spike antibodies (3525, 0.2 μg/mL) in the brain of the COVID-19 patient confirmed by PCR. (Courtesy of Dr. Nuovo Gerard J., OSU)
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<strong>Figure 12 Overexpression Validation in Spike Transfected 293 Cells </strong><br> Loading: 15 μg per lane of 293 cell lysate. Antibodies: SARS-CoV-2 (COVID-19) Spike, 3525 (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
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<strong>Figure 13 Immunofluorescence Validation of SARS-CoV-2 (COVID-19) Spike in 293T Cells</strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed 293T cells labeling SARS-CoV-2 (COVID-19) Spike with 3525 at 1 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).
14 / 14
<strong>Figure 14 ELISA Test</strong><br> Antibodies: SARS-CoV-2 (COVID-19) Spike antibody, 3525 (1 μg/mL). A direct ELISA was performed using immunogen or control peptide as coating antigen and the anti-SARS-CoV-2 (COVID-19) Spike antibody as the capture antibody. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:20000 dilution. Detection range is from 0.5 ng/mL to 1000 ng/mL.
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<strong>Figure 1 Immunofluorescent Validation of 3525 in SARS-CoV-2 Infected Lung Tissue (Singh et al., Nature Microbiology, 2021) </strong><br>Multilabel confocal immunofluorescence microscopy of formalin-fixed paraffin-embedded lung sections from rhesus macaques infected with SARS-CoV-2. SARS-CoV-2 spike-specific antibodies, 3525 ProSci Inc. (k, n) (turquoise); Ki67 (magenta) and neutrophil marker CD66abce (yellow) (k); pan-macrophage marker CD68 (magenta) (n) and DAPI (blue).
<strong> Figure 2 Immunofluorescent Validation of 3525 in SARS-CoV-2 Infected Nose and Tonsil (Singh et al., Nature Microbiology, 2021) </strong><br> Multi-label confocal immunofluorescence microscopy of nasal epithelium (20X-b, 63xh) and tonsil (20X-c,63X-i) from rhesus macaques infected with SARS-CoV-2 with SARS-CoV-2 spike-specific antibodies, 3525 ProSci Inc. (turquoise), DAPI (blue). Rabbit IgG isotype control antibody was used to stain the tissues to rule out any non-specific staining (e, f).
<strong> Figure 3 Immunofluorescent Validation of 3525 in SARS-CoV-2 Infected Lung Tissue (Singh et al., Nature Microbiology, 2021) </strong><br> Multilabel confocal immunofluorescence microscopy of formalin-fixed paraffin-embedded lung sections from rhesus macaques infected with SARS-CoV-2. SARS-CoV-2 spike-specific antibodies, 3525 ProSci Inc. (k-s) (turquoise); Ki67 (magenta) and neutrophil marker CD66abce (yellow) (k-m); pan-macrophage marker CD68 (magenta) (n-p); HLA-DR (magenta) and pDC marker CD123 (yellow) (q?s) and DAPI (blue).
<strong>Figure 4 Immunohistochemistry Validation of SARS-CoV-2 (COVID-19) Spike 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 S2 antibody (3525, 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 spike protein signal was observed in macrophages and airway epithelium of COVID-19 patient lung, but not in non-COVID-19 patient lung.
<strong>Figure 5 Immunohistochemistry Validation of SARS-CoV-2 (COVID-19) Spike 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 S2 antibody (3525, 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 spike protein signal was observed in macrophages of COVID-19 patient lung.
<strong>Figure 6 IHC/IF Validation in COVID-19 Patient Sample </strong><br> (Nuovo et al., 2020) Detection of SARS-CoV-2 proteins in nasopharyngeal swab cell preparations. B. An intense signal for covid-19 spike protein <strong> tested by SARS-CoV-2 spike antibodies (3525) </strong> was observed in the glandular cells. F-H. Co-expression of spike detected by spike antibodies (3525, 0.2 μg/mL) and envelope proteins detected by envelope antibodies (3531, 2 μg/mL) of SARS-CoV-2 (F) documented localization of each protein to glandular cells (G, yellow). No signal was seen in oral swabs of positive cases (H). <strong>Both the spike and envelope protein detected by anti-spike antibodies (3525) and anti-envelope antibodies (3531) produced a signal in the nasopharyngeal swabs of the three cases and no signal was evident in the nasopharyngeal swabs of the seven controls. </strong>
<strong>Figure 7 IF Validation of SARS-CoV2 Spike in COVID-19 Patient Lung </strong><br> (Magro et al., 2020) SARS-CoV2 spike protein (red, panel C) detected detected by anti-spike antibodies (3525, 0.2 μg/mL) colocalized with C4d (green in panel d, merged in yellow). Spike protein (red, panel g) was also colocalized with C5b-9 (green in panel f&h, merged in yellow)
<strong>Figure 8 IF Validation of SARS-CoV2 Spike in COVID-19 Patient Skin</strong><br> (Magro et al., 2020) C4d is highlighted green while COVID-19 spike protein detected by anti-spike antibodies (3525, 0.2 μg/mL) shows a red staining pattern; a yellow signal is discernible indicative of co-localization of C4d and viral protein within the microvasculature.
<strong>Figure 9 IHC Validation of SARS-CoV2 Spike in the Nasopharyngeal Swab Sample of the COVID-19 Patient </strong><br> Strong spike signal was detected by anti-spike antibodies (3525, 0.2 μg/mL) in the nasopharyngeal swab sample of the COVID-19 patient and no spike signal was observed in the sample of the COVID-19 negative patient. COVID-19 cases were confirmed by PCR. (Courtesy of Dr. Nuovo Gerard J., OSU)
<strong>Figure 10 IHC Validation of SARS-CoV2 Spike in COVID-19 Patient Lung</strong><br> Strong spike signal was detected by anti-spike antibodies (3525, 0.2 μg/mL) in the lung of the COVID-19 patient confirmed by PCR. (Courtesy of Dr. Nuovo Gerard J., OSU)
<strong>Figure 11 IHC Validation of SARS-CoV2 Spike in COVID-19 Patient Brain</strong><br> Spike protein was detected by anti-spike antibodies (3525, 0.2 μg/mL) in the brain of the COVID-19 patient confirmed by PCR. (Courtesy of Dr. Nuovo Gerard J., OSU)
<strong>Figure 12 Overexpression Validation in Spike Transfected 293 Cells </strong><br> Loading: 15 μg per lane of 293 cell lysate. Antibodies: SARS-CoV-2 (COVID-19) Spike, 3525 (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
<strong>Figure 13 Immunofluorescence Validation of SARS-CoV-2 (COVID-19) Spike in 293T Cells</strong><br> Immunofluorescent analysis of 4% paraformaldehyde-fixed 293T cells labeling SARS-CoV-2 (COVID-19) Spike with 3525 at 1 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue).
<strong>Figure 14 ELISA Test</strong><br> Antibodies: SARS-CoV-2 (COVID-19) Spike antibody, 3525 (1 μg/mL). A direct ELISA was performed using immunogen or control peptide as coating antigen and the anti-SARS-CoV-2 (COVID-19) Spike antibody as the capture antibody. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:20000 dilution. Detection range is from 0.5 ng/mL to 1000 ng/mL.

Documents

Further Information

Additional Names:
SARS-CoV-2 (COVID-19) Spike Antibody: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), Surface Glycoprotein, Spike protein
Application Note:
WB: 1 μg/mL; IF: 1 μg/mL. IHC: 0.2 μg/mL

Antibody validated: Immunofluorescence and Western blot in human samples. Immunohisochemistry and immunofuorescence in COVID-19 patient samples. It will detect 4 ng of free peptide at 1 μg/mL. The immunogen for this is within the last 50 aa of the spike protein - a peptide corresponding to 20 amino acids near the carboxy terminus of SARS-CoV-2 (COVID-19) Spike glycoprotein. The Extracellular domain (ECD) is from aa 1 to 1208 (full length 1273aa). Therefore, this antibody detects the transmembrane and cytoplasm domains at the C terminus, but does not detect the ECD (which is the region expressed in many commercially available spike proteins). 3525 can be used for the detection of full length spike protein and spike protein in COVID-19 patient samples. 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 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: (100%)
Immunogen:
Anti-SARS-CoV-2 (COVID-19) Spike antibody (3525) was raised against a peptide corresponding to 20 amino acids near the carboxy terminus of SARS-CoV-2 (COVID-19) Spike glycoprotein.

The immunogen is located within the last 50 amino acids of SARS-CoV-2 (COVID-19) Spike protein.
ISOFORMS:
SARS-CoV-2 (COVID-19) Spike has one isoform (1273aa).
NCBI Gene ID #:
43740568
NCBI Official Name:
surface glycoprotein
NCBI Official Symbol:
S
NCBI Organism:
Wuhan seafood market pneumonia virus
Physical State:
Liquid
Protein Accession #:
QHD43416
Protein GI Number:
1791269090
Purification:
SARS-CoV-2 (COVID-19) Spike 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

  1. Nuovo GJ et al. Cytologic and molecular correlates of?SARS-CoV-2 infection of the nasopharynx. Ann Diagn Pathol. 2020; 48:151565. doi: 10.1016/j.anndiagpath.2020.151565. PMID: 32659620
  2. Magro C, Mulvey JJ, Berlin D, Nuovo G, Salvatore S, Harp J, Baxter-Stoltzfus A, Laurence J. Complement associated microvascular injury and thrombosis in the pathogenesis of severe COVID-19 infection: A report of five cases. Transl Res. 2020; 220:1-13. doi: 10.1016/j.trsl.2020.04.007. PMID: 32299776
  3. Ko, et al. Discordant anti-SARS-CoV-2 spike protein and RNA staining in cutaneous perniotic lesions suggests endothelial deposition of cleaved spike protein. J Cutan Pathol . 2021 Jan;48(1):47-52. doi: 10.1111/cup.13866. Epub 2020 Oct 1.PMID: 32895985?
  4. Magro, et al. Docked severe acute respiratory syndrome coronavirus 2 proteins within the cutaneous and subcutaneous microvasculature and their role in the pathogenesis of severe coronavirus disease 2019. Hum Pathol. 2020 Dec;106:106-116. doi: 10.1016/j.humpath.2020.10.002. Epub 2020 Oct 12.PMID: 33058948
  5. Magro, et al. The differing pathophysiologies that underlie COVID-19-associated perniosis and thrombotic retiform purpura: a case series. Br J Dermatol. 2020 Jul 22;10.1111/bjd.19415. doi: 10.1111/bjd.19415PMID: 32779733
  6. Mulvey, et al. Analysis of complement deposition and viral RNA in placentas of COVID-19 patients. Ann Diagn Pathol. 2020 Jun;46:151530. doi: 10.1016/j.anndiagpath.2020.151530. Epub 2020 Apr 25.PMID: 32387855
  7. Nuovo, et al. Strong homology between SARS-CoV-2 envelope protein and a Mycobacterium sp. antigen allows rapid diagnosis of Mycobacterial infections and may provide specific anti-SARS-CoV-2 immunity via the BCG vaccine. Ann Diagn Pathol. 2020 Oct;48:151600. doi: 10.1016/j.anndiagpath.2020.151600. Epub 2020 Aug 13.PMID: 32805515
  8. Singh, et al. Responses to acute infection with SARS-CoV-2 in the lungs of rhesus macaques, baboons and marmosets. Nat Microbiol. 2021 Jan;6(1):73-86. doi: 10.1038/s41564-020-00841-4. Epub 2020 Dec 18PMID: 33340034

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