ORF8 Antibody (1041422) [Unconjugated]

Western blot shows recombinant SARS-CoV-2 ORF8 protein. PVDF membrane was probed with 2 µg/mL of Mouse Anti-SARS-CoV-2 ORF8 Monoclonal Antibody (Catalog # MAB10820) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for ORF8 at approximately 14 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

ORF8 was detected in immersion fixed HEK293 human embryonic kidney cell line transfected with SARS-CoV-2 (positive staining) and HEK293 human embryonic kidney cell line (non-transfected, negative staining) using Mouse Anti-SARS-CoV-2 ORF8 Monoclonal Antibody (Catalog # MAB10820) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. Staining was performed using our protocol for Fluorescent ICC Staining of Non-adherent Cells.

ORF8 was detected in immersion fixed paraffin-embedded sections of SARS-CoV-2 infected human lung using Mouse Anti-SARS-CoV-2 ORF8 Monoclonal Antibody (Catalog # MAB10820) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (VC001). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to immunoreactive profiles scattered throughout the tissue. Staining was performed using our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

• Reactivity: V
• Applications: WB, IHC, ICC/IF
• Clone: 1041422
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated

ORF8 Antibody (1041422) [Unconjugated] Summary

• Immunogen: E. coli-derived human SARS-CoV-2 ORF8
  Met16-His121
  Accession # YP_009724369
• Specificity: Detects human SARS-CoV-2 ORF8 in direct ELISAs.
• Source: N/A
• Isotype: IgG2a
• Clonality: Monoclonal
• Host: Mouse
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 8-25 ug/mL
  • Immunohistochemistry 5-25 ug/mL
  • Western Blot 2 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for ORF8 Antibody (1041422) [Unconjugated]

• ORF8

Background

SARS-CoV-2, which causes the global pandemic coronavirus disease 2019 (Covid-19), belongs to a family of viruses known as coronaviruses that are commonly comprised of four structural proteins: Spike protein(S), Envelope protein (E), Membrane protein (M), and Nucleocapsid protein (N) (1). M protein is the most abundant structural protein in the coronavirus membrane, and it is predicted to span the membrane three times, with a short N-terminal domain outside the viral envelope and a long C-terminal domain inside the virion (2). SARS-CoV-2 M protein is a 222 amino acid (aa) glycoprotein that is composed of an 18 aa N-terminal domain on the viral surface, 3 transmembrane domains, and a 122 aa C-terminal domain inside the viral envelope. SARS-CoV-2 M protein shares 89.14%, 98.6%, 98.2%, and 38.36% aa similarity with SARS-CoV-1, bat SARS-CoV, pangolin SARS-CoV, and MERS-CoV M proteins, respectively (3). The M protein of coronavirus plays an important role in assembly of viral particles by interacting with other structural proteins, especially with the E protein (4, 5). In SARS-CoV-2, M protein, combined with E protein, regulates intracellular trafficking of the S Protein and its unique furin-mediated processing (6).

1. Wu, F. et al. (2020) Nature 579:265.
2. Mousavizadeh, L. and S. Ghasemi (2020) J. Microbiol. Immunol. Infect. doi:10.1016/j.jmii.2020.03.022.
3. Thomas, S. (2020) Pathog. Immun. 5:342.
4. Masters, P.S. (2006) Adv. Virus. Res. 66:193.
5. Corse, E. and C.E. Machamer (2003) Virology 312:25.
6. Boson, B. et al. (2020) bioRxiv doi:10.1101/2020.08.24.260901.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for ORF8 Antibody (MAB10820)(1)

Publication using MAB10820

• Melano, I;Azamor, T;Caetano, CC;Meyer, NM;Onwubueke, C;Visperas, A;Familiar-Macedo, D;Salem, GM;Soos, BL;Calabrese, CM;Choi, YJ;Chen, S;Choi, Y;Wu, X;Vasconcelos, Z;Comhair, SA;Nielsen-Saines, K;Calabrese, LH;Husni, ME;Jung, JU;Piuzzi, NS;Foo, SS;Chen, W; SARS-CoV-2 ORF8 drives osteoclastogenesis in preexisting immune-mediated inflammatory diseases JCI insight 2024-12-20 [PMID: 39704167] (ELISA Capture, Human)

Bioinformatics

• Uniprot:
  • YP_009724369()