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SARS-CoV-2 NSP16 Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB10940

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MAB10940-100
MAB10940-SP

Key Product Details

Species Reactivity

SARS-CoV-2

Applications

Western Blot, ELISA, Simple Western

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2B Clone # 1044311

Product Specifications

Immunogen

E. coli-derived SARS-CoV-2 NSP16/NSP10
SARS-CoV-2 NSP16 (Ser1-Asn298) and SARS-CoV-2 NSP10 (Ala1-Gln139)
Accession # YP_009725311.1 and YP_009725306.1

Specificity

Detects SARS-CoV-2 NSP16/NSP10 in direct ELISAs

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2B

Scientific Data Images for SARS-CoV-2 NSP16 Antibody

Detection of SARS-CoV-2 NSP16 by Western Blot.

Western blot shows recombinant SARS-CoV-2 NSP16/NSP10. PVDF membrane was probed with 1 µg/mL of Mouse Anti-SARS-CoV-2 NSP16 Monoclonal Antibody (Catalog # MAB10940) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for NSP16/NSP10 at approximately 34 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Detection of SARS-CoV-2 NSP16 by Simple WesternTM.

Simple Western lane view shows recombinant SARS-CoV-2 Nsp10/Nsp16, loaded at 0.2 mg/mL. A specific band was detected for NSP16 at approximately 39 kDa (as indicated) using 20 µg/mL of Mouse Anti-SARS-CoV-2 NSP16 Monoclonal Antibody (Catalog # MAB10940) . This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

SARS-CoV-2 NSP16/NSP10 ELISA Standard Curve.

Recombinant SARS-CoV-2 NSP16/NSP10 protein was serially diluted 2-fold and captured by Mouse Anti-SARS-CoV-2 His-tag Complex Monoclonal Antibody (Catalog # MAB10941) coated on a Clear Polystyrene Microplate (Catalog # DY990). Mouse Anti-SARS-CoV-2 NSP16 Monoclonal Antibody (Catalog # MAB10940) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998) followed by Substrate Solution (Catalog # DY999) and stopping the enzymatic reaction with Stop Solution (Catalog # DY994).

Applications for SARS-CoV-2 NSP16 Antibody

Application
Recommended Usage

ELISA

This antibody functions as an ELISA detection antibody when paired with Mouse Anti-SARS-CoV-2 NSP16 Monoclonal Antibody (Catalog # MAB10941).This product is intended for assay development on various assay platforms requiring antibody pairs.

Simple Western

20 µg/mL
Sample: Recombinant SARS-CoV-2 Nsp10/Nsp16

Western Blot

1 µg/mL
Sample: Recombinant SARS-CoV-2 NSP16/NSP10

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

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Formulation

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.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & 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.

Background: NSP16

Non-structural protein 16 (NSP16) and NSP10 are two 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. NSP16 is a small monomeric 298 amino acid protein with a characteristic fold of the class I methyltransferase (MTase) family with a seven-stranded B-sheet surrounded by alpha-helices and loops that bind the cap and form a cap-binding groove and S-adenosylmethionine (SAM) cleft (2, 3). NSP10 is a small 139 amino acid protein capable of forming independent dodecameric structures composed of four identical trimers where each monomer is capable of binding two zinc ions (4, 5). Both the NSP16 and NSP10 SARS-CoV2 sequences are highly conserved across coronaviruses (3). NSP16 catalyzes 2'-O-MTase activity specifically on capped N7-methylated RNA through methylation of the 2'-hydroxy group of adenine using SAM as a methyl donor (6). The viral RNA cap structure protects it from degradation, promotes mRNA translation, and prevents the viral RNA from being recognized by innate immune mechanisms (7). NSP16 was shown to interact strongly with NSP10 (8) in a monomeric form via hydrophic interactions and hydrogen bonds in the interface (3). The MTase activity of NSP16 requires interaction with NSP10 (2, 7). NSP10 interaction extends and narrows the RNA-binding groove that accommodates the substrate and stabilizes NSP16 (2, 3, 9). The dimerization interface between NSP16 and NSP10 is critical and can be used as a target to effectively reduce replication and pathogenesis (10, 11).

References

  1. Snijder, E.J. et al. (2016) Adv. Virus Res 96:59.
  2. Chen, Y. et al. (2011) PLoS Pathog. 7:e1002294.
  3. Rosas-Lemus, M. et al. (2020) bioRxiv In press.
  4. Matthes, N. et al. (2006) FEBS Lett. 580:4143.
  5. Su, D. et al. (2006) J. Virol. 80:7902.
  6. Bouvet, M. et al. (2012) Proc. Natl. Acad. Sci. U.S.A. 109:9372.
  7. Decroly, E. et al. (2011) PLoS Pathog. 7:e1002059.
  8. Pan, J. et al. (2008) PLoS One 3:e3299.
  9. Bouvet, M. et al. (2014) J. Biol. Chem. 289:25783.
  10. Ma, Y. et al. (2015) Proc. Natl. Acad. Sci. 112:9436.
  11. Wang, Y. et al. (2015) J. Virol 89:8416.

Long Name

Non-Structural Protein 16

Additional NSP16 Products

Product Documents for SARS-CoV-2 NSP16 Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for SARS-CoV-2 NSP16 Antibody

For research use only

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