SARS-CoV NSP8 Antibody (1044736) [Unconjugated]

Western blot shows recombinant SARS-CoV-2 Nsp8 protein. PVDF membrane was probed with 1 µg/mL of Mouse Anti-SARS-CoV-2 Nsp8 Monoclonal Antibody (Catalog # MAB12502) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for Nsp8 at approximately 24 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Simple Western lane view shows recombinant SARS-CoV-2 Nsp8 protein, loaded at 0.2 mg/mL. A specific band was detected for Nsp8 at approximately 29 kDa (as indicated) using 20 µg/mL of Mouse Anti-SARS-CoV-2 Membrane Monoclonal Antibody (Catalog # MAB12502) . This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

• Reactivity: V
• Applications: WB, Simple Western
• Clone: 1044736
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated

SARS-CoV NSP8 Antibody (1044736) [Unconjugated] Summary

• Immunogen: E. coli-derived SARS-CoV-2 NSP8 protein
  Ala1-Gln198
  Accession # YP_009725304.1
• Specificity: Detects SARS-CoV-2 NSP8 in direct ELISAs.
• Source: N/A
• Isotype: IgG2b
• Clonality: Monoclonal
• Host: Mouse
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • Simple Western 20 ug/mL
  • Western Blot 1 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 SARS-CoV NSP8 Antibody (1044736) [Unconjugated]

• COVID-19
• NSP8

Background

Non-structural protein 8 (NSP8) is one of several functional proteins released by ORF1a-encoded protease cleavage of the pp1a and pp1ab replicase polyproteins expressed from the coronavirus (CoV) genome (1). The NSPs are involved in the replication and transcription of the viral RNA and not incorporated within the virion particles. Coronaviruses include various highly pathogenic strains such as SARS-CoV, MERS-CoV and SARS-CoV2 that have had significant impact on humans as well as strains that have negatively impacted livestock. NSP8 is a small 198 amino acid protein that forms a dimer with NSP7 and subsequently assembles into a large hexadecameric structure (2). The NSP8 sequence is highly conserved across coronaviruses (2). The NSP8 monomers in each of two asymmetric units can adopt two different conformations: a "golf-club like" structure with the N-terminal shaft and C-terminal head or a bent N-terminal shaft and C-terminal head domain (2). The units stack to form a supercomplex like bricks with layers of NSP7 filling the spaces in between. The supercomplexes are stacked to form a channel with electrostatic properties that would allow RNA to pass through the channel, likely to facilitate efficient replication and transcription. NSP8 was also shown to be able to polymerize small oligomers in a sequence-specific fashion and was consequently proposed to act as an RNA primase for the viral RNA-dependent RNA polymerase (RdRp), NSP12, in SARS-CoV (3). In SARS-CoV- 2, RdRp has been shown to have little activity without NSP8/7 acting as cofactors to form a complex (4) making NSP8 critical for viral polymerase activity. NSP8 was shown to interact with several other viral NSP proteins, including NSP2 and NSP9, (5) as well as multiple host cell proteins (6). NSP8 showed interaction with 3 different signal recognition particle (SRP) host cell proteins suggesting the virus hijacks the SEC61-mediated protein translocation pathway for entry into the endoplasmic reticulum and NSP8 may be involved in targeting (6). 

1. Snijder, E.J. et al. (2016) Adv. Virus Res. 96:59.
2. Zhai, Y. et al. (2005) Nat. Struct. Mol. Bio. 12:980.
3. Subissi, L. et al. (2014) Antiviral Res. 101:122.
4. Yin, W. et al. (2020) Science 368:1499.
5. von Brunn, A. et al. (2007) PLoS One 2:e459.
6. Gordon, D.E. et al. (2020) Nature 583:459.

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 SARS-CoV NSP8 Antibody (MAB12502)(1)

Publication using MAB12502

• Duan, T;Xing, C;Chu, J;Deng, X;Du, Y;Liu, X;Hu, Y;Qian, C;Yin, B;Wang, HY;Wang, RF; ACE2-dependent and -independent SARS-CoV-2 entries dictate viral replication and inflammatory response during infection Nature cell biology 2024-03-21 [PMID: 38514841] (Western Blot, Human)

Bioinformatics

• Uniprot:
  • YP_009725304.1()