Key Product Details

Species Reactivity

SARS-CoV-2

Applications

Immunohistochemistry, ELISA, Immunocytochemistry, Simple Western

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # 1045410
View all NSP1 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived Sars-CoV-2 NSP1 protein
Met1-Gly180
Accession # YP_009725297.1

Specificity

Detects Sars-CoV-2 NSP1 protein in ELISA.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Scientific Data Images for SARS-CoV-2 NSP1 Antibody

NSP1 in HEK293 Human Cell Line Transfected with SARS-CoV-2.

NSP1 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 NSP1 Monoclonal Antibody (Catalog # MAB10980) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

NSP1 in SARS-CoV-2 infected human lung.

NSP1 was detected in immersion fixed paraffin-embedded sections of SARS-CoV-2 infected human lung using Mouse Anti-SARS-CoV-2 NSP1 Monoclonal Antibody (Catalog # MAB10980) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to immunoreactive profiles scattered throughout the tissue. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Detection of SARS-CoV-2 NSP1 by Simple WesternTM.

Simple Western lane view shows recombinant SARS-CoV-2 Nsp1, loaded at 0.2 mg/mL. A specific band was detected for NSP1 at approximately 28 kDa (as indicated) using 20 µg/mL of Mouse Anti-SARS-CoV-2 NSP1 Monoclonal Antibody (Catalog # MAB10980). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

SARS-CoV-2 NSP1 ELISA Standard Curve.

Recombinant SARS-CoV-2 NSP1 protein was serially diluted 2-fold and captured by Mouse Anti-SARS-CoV-2 NSP1 Monoclonal Antibody (Catalog # MAB10980) coated on a Clear Polystyrene Microplate (Catalog # DY990). Mouse Anti-SARS-CoV-2 NSP1 Monoclonal Antibody (Catalog # MAB10991) 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 NSP1 Antibody

Application
Recommended Usage

ELISA

This antibody functions as an ELISA capture antibody when paired with Mouse Anti-SARS-CoV-2 NSP1 Monoclonal Antibody (Catalog # MAB10991).

This product is intended for assay development on various assay platforms requiring antibody pairs.

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed HEK293 human embryonic kidney cell line transfected with SARS-CoV-2

Immunohistochemistry

5-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of SARS-CoV-2 infected human lung

Simple Western

20 µg/mL
Sample: Recombinant SARS-CoV-2 Nsp1

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.

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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: NSP1

Non-structural protein 1 (NSP1) 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. NSP1, also known as the host shutoff factor, is a small 180 amino acid highly conserved protein among the first to be expressed following cell entry.  It is composed of an N-terminal domain, a linker region, and a C-terminal domain with a conserved KH motif that is required and sufficient for specific contacts with ribosomes (2, 3). The C-terminal domain binds tightly to and sterically occludes the entrance region of the mRNA channel in free 40S subunits, 43S pre-initiation complex, and empty, non-translating 80S ribosomes (2, 3) to inhibit translation. NSP1 blocks the innate immune response through suppression of host gene expression. By preventing translation of interferon and downstream signaling responses, it plays a key role in immune evasion (2, 4, 5). NSP1 also induces endonucleolytic cleavage of host mRNAs (6,7). Concomitant translation of more efficiently recognized untranslated regions (UTR) of viral mRNA along with resistance to endonucleolytic cleavage (7) is thought to lead to a switch to production of viral mRNA over host cell mRNA during an infection (3,8). Given the critical role NSP1 plays in virulence, it is an attractive target for small-molecule inhibition and vaccination development (9, 10).  

References

  1. Snijder, E.J. et al. (2016) Adv. Virus Res 96:59.
  2. Thoms, M. et al. (2020) Science 369:1249.
  3. Schubert, K. et al. (2020) Nat. Struct. Mol. Biol. 27:959.
  4. Narayanan, K. et al. (2008) J. Virol. 82:4471.
  5. Jauregui, A.R. et al. (2013) PLoS One. 8:e62416.
  6. Kamitani, W. et al. (2006) Proc. Natl. Acad. Sci. USA 103:12885.
  7. Huang, C. et al. (2011) PLoS Pathog. 7:e1002433.
  8. Hartenian, E. et al. (2020) J. Biol. Chem. 295:12910. 
  9. Zust, R. et al. (2007) PLoS Pathog. 3:e109.
  10. De Lima Menezes, G. and R.A. da Silva (2020) J. Biomol. Struct. Dyn. (In press). 

Long Name

Non-Structural Protein 1

UniProt

YP_009725297.1

Additional NSP1 Products

Product Documents for SARS-CoV-2 NSP1 Antibody

Certificate of Analysis

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Product Specific Notices for SARS-CoV-2 NSP1 Antibody

For research use only

Related Research Areas

Infectious Diseases

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Protocols

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