Recombinant SARS-CoV-2 NSP16/NSP10-His Complex, CF

Name: Recombinant SARS-CoV-2 NSP16/NSP10-His Complex, CF
Brand: R&D Systems
SKU: 10634-CV
Rating: 4 (1 reviews)

Catalog # 10634-CV

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Key Product Details

Source:	E. coli
Accession #:	YP_009725311.1 (NSP16) and YP_009725306.1 (NSP10)

View Full Technical Details for Recombinant SARS-CoV-2 NSP16/NSP10-His Complex, CF

Product Specifications

Source

E. coli-derived sars-cov-2 NSP16/NSP10 protein

SARS-CoV-2 NSP16 (Ser1-Asn298) Accession # YP_009725311.1
SARS-CoV-2 NSP10 (Ala1-Gln139) Accession # YP_009725306.1	6-His tag
N-terminus	C-terminus

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin level

<1.0 EU per 1 μg of the protein by the LAL method.

N-terminal sequence Analysis

Ser1 (NSP16) & Ala1 (NSP10)

Predicted Molecular Mass

33 kDa (NSP16) & 16 kDa (NSP10)

SDS-PAGE

34 kDa (NSP16) & 17 kDa (NSP10), under reducing conditions

Recombinant SARS-CoV-2 NSP16/NSP10-His Complex, CF Scientific Data Examples

	Recombinant SARS-CoV-2 NSP16/NSP10-His Complex SDS-PAGE. 2 μg/lane of Recombinant SARS-CoV-2 NSP16/NSP10-His Complex (Catalog # 10634-CV) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 34 kDa and 17 kDa under reducing conditions.

Formulation, Preparation and Storage

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

10634-CV

Formulation	Supplied as a 0.2 μm filtered solution in Tris, NaCl, TCEP and Glycerol.
Shipping	The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage:	Use a manual defrost freezer and avoid repeated freeze-thaw cycles. 6 months from date of receipt, -20 to -70 °C as supplied. 3 months, -20 to -70 °C under sterile conditions after opening.

Reconstitution Calculator

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: NSP16/NSP10

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).

Item	Value
References	Snijder, E.J. et al. (2016) Adv. Virus Res 96:59. Chen, Y. et al. (2011) PLoS Pathog. 7:e1002294. Rosas-Lemus, M. et al. (2020) bioRxiv In press. Matthes, N. et al. (2006) FEBS Lett. 580:4143. Su, D. et al. (2006) J. Virol. 80:7902. Bouvet, M. et al. (2012) Proc. Natl. Acad. Sci. U.S.A. 109:9372. Decroly, E. et al. (2011) PLoS Pathog. 7:e1002059. Pan, J. et al. (2008) PLoS One 3:e3299. Bouvet, M. et al. (2014) J. Biol. Chem. 289:25783. Ma, Y. et al. (2015) Proc. Natl. Acad. Sci. 112:9436. Wang, Y. et al. (2015) J. Virol 89:8416.
Long Name	NSP16/NSP10
Alternate Names	NSP16/NSP10

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Recombinant SARS-CoV-2 NSP16/NSP10-His Complex, CF

Name: Anonymous

Application: In vitro bioactivity in cell culture

Reason for Rating: Stimulated immune cells evidenced by increased cytokine expression.

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Recombinant SARS-CoV-2 NSP16/NSP10-His Complex, CF

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Recombinant SARS-CoV-2 NSP16/NSP10-His Complex, CF Scientific Data Examples

Formulation, Preparation and Storage

Carrier Free

Reconstitution Calculator

Background: NSP16/NSP10

Citations for Recombinant SARS-CoV-2 NSP16/NSP10-His Complex, CF

Customer Reviews (1)

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Recombinant SARS-CoV-2 NSP16/NSP10-His Complex, CF

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