SARS-CoV Nucleocapsid Protein E. coli

Cat. No.	Size	Price
RD972052100	0.1 mg

Technical Data

Type

Recombinant protein

Description

Total 437 AA. MW: 47.8 kDa (calculated). UniProtKB acc.no. P59595. N-Terminal His-tag 16 AA.

Amino Acid Sequence

MRGSHHHHHHGMASHMSDNGPQSNQRSAPRITFGGPTDSTDNNQNGGRNGARPKQRRPQGLPNNTASWFTALTQHGKEELRFPRGQGVPINTNSGPDDQIGYYRRATRRVRGGDGKMKELSPRWYFYYLGTGPEASLPYGANKEGIVWVATEGALNTPKDHIGTRNPNNNAATVLQLPQGTTLPKGFYAEGSRGGSQASSRSSSRSRGNSRNSTPGSSRGNSPARMASGGGETALALLLLDRLNQLESKVSGKGQQQQGQTVTKKSAAEASKKPRQKRTATKQYNVTQAFGRRGPEQTQGNFGDQDLIRQGTDYKHWPQIAQFAPSASAFFGMSRIGMEVTPSGTWLTYHGAIKLDDKDPQFKDNVILLNKHIDAYKTFPPTEPKKDKKKKTDEAQPLPQRQKKQPTVTLLPAADMDDFSRQLQNSMSGASADSTQA

Source

E. coli

Purity

˃ 90 % by SDS-PAGE

SDS-PAGE Gel

12% SDS-PAGE separation of SARS-CoV Nucleocapsid protein
1. M.W. marker – 14, 21, 31, 45, 66, 97 kDa
2. reduced and heated sample, 5μg/lane
3. non-reduced and non-heated sample, 5μg/lane

Endotoxin

< 1.0 EU/µg

Formulation

Filtered (0,4 μm) and lyophilized in 0.5 mg/mL in 0.05 M Acetate buffer pH= 4.0

Reconstitution

Add 0.1M Acetate buffer pH4 to prepare a working stock solution of approximately 0.5 mg/mL and let the lyophilized pellet dissolve completely. For conversion into higher pH value, we recommend intensive dilution by relevant buffer to a concentration of 10μg/mL. In higher concentrations the solubility of this antigen is limited.

Applications

Western blotting

Shipping

At ambient temperature. Upon receipt, store the product at the temperature recommended below.

Storage/Expiration

Store the lyophilized protein at -80 °C. Lyophilized protein remains stable until the expiry date when stored at -80 °C. Aliquot reconstituted protein to avoid repeated freezing/thawing cycles and store at -80 °C for long term storage. Reconstituted protein can be stored at 4 °C for a week.

Quality Control Test

BCA to determine quantity of the protein.
SDS PAGE to determine purity of the protein. LAL to determine quantity of endotoxin.

Note

This product is intended for research use only.

Summary

Research topic

Immune Response, Infection and Inflammation, COVID-19

Summary

Severe acute respiratory syndrome (SARS) coronavirus (SARS-CoV) caused a severe outbreak in several regions of the world in 2003. The virus is a novel coronavirus isolated from patients exhibiting atypical pneumonia and may have originated from wild animals such as civet cats in southern China. The genome of SARS-CoV is a positive-sense, single-stranded RNA whose sequence is distantly related to all known coronaviruses that infect humans and animals. Like other known coronaviruses, SARS-CoV is an enveloped virus containing three outer structural proteins, namely the membrane (M), envelope (E), and spike (S) proteins. The nucleocapsid (N) protein together with the viral RNA genome form a helical core located within the viral envelope.

The severe acute respiratory syndrome coronavirus nucleocapsid protein (SARS-CoV NP), a phosphoprotein of 48 kDa, is the most abundant protein in the virus-infected cells. Its primary function is to package the ∼30 kb single stranded, 5′-capped positive strand viral genome RNA molecule into a ribonucleoprotein (RNP) complex called the capsid. Ribonucleocapsid packaging is a fundamental part of viral self-assembly and the RNP complex constitutes the essential template for replication by the RNA-dependent RNA polymerase complex. In addition, the N-protein of the SARS-CoV has been shown to modulate the host cellular machinery and may serve regulatory roles during its viral life cycle.

As the most abundantly expressed structural protein during infection, SARS-CoV NP is highly detectable in SARS patients. Therefore, this protein may serve as one of the immunodominant antigens in the early diagnosis of infection. Furthermore, researchers have suggested that antibody against the N protein could modulate cytokine responses such as IL-11; non-neutralizing antibodies against N protein were found to protect mice against lethal infection.

Product References (1)

References

Shin GC, Chung YS, Kim IS, Cho HW, Kang C. Preparation and characterization of a novel monoclonal antibody specific to severe acute respiratory syndrome-coronavirus nucleocapsid protein. Virus Res. 2006 Dec;122(1-2):109-18. doi: 10.1016/j.virusres.2006.07.004. Epub 2006 Aug 30. PubMed PMID: 16942813. PubMed CentralPMCID: PMC7114302. See more on PubMed

Summary References (18)

References to SARS-CoV

Di B, Hao W, Gao Y, Wang M, Wang YD, Qiu LW, Wen K, Zhou DH, Wu XW, Lu EJ,Liao ZY, Mei YB, Zheng BJ, Che XY. Monoclonal antibody-based antigen captureenzyme-linked immunosorbent assay reveals high sensitivity of the nucleocapsidprotein in acute-phase sera of severe acute respiratory syndrome patients. ClinDiagn Lab Immunol. 2005 Jan;12(1):135-40. PubMed PMID: 15642998; PubMed CentralPMCID: PMC540218. See more on PubMed
He R, Dobie F, Ballantine M, Leeson A, Li Y, Bastien N, Cutts T, Andonov A,Cao J, Booth TF, Plummer FA, Tyler S, Baker L, Li X. Analysis of multimerization of the SARS coronavirus nucleocapsid protein. Biochem Biophys Res Commun. 2004Apr 2;316(2):476-83. PubMed PMID: 15020242. See more on PubMed
Hsieh PK, Chang SC, Huang CC, Lee TT, Hsiao CW, Kou YH, Chen IY, Chang CK,Huang TH, Chang MF. Assembly of severe acute respiratory syndrome coronavirus RNApackaging signal into virus-like particles is nucleocapsid dependent. J Virol.2005 Nov;79(22):13848-55. PubMed PMID: 16254320; PubMed Central PMCID:PMC1280188. See more on PubMed
Huang LR, Chiu CM, Yeh SH, Huang WH, Hsueh PR, Yang WZ, Yang JY, Su IJ, Chang SC, Chen PJ. Evaluation of antibody responses against SARS coronaviralnucleocapsid or spike proteins by immunoblotting or ELISA. J Med Virol. 2004Jul;73(3):338-46. PubMed PMID: 15170626. See more on PubMed
Chang CK, Hou MH, Chang CF, Hsiao CD, Huang TH. The SARS coronavirusnucleocapsid protein--forms and functions. Antiviral Res. 2014 Mar;103:39-50.doi: 10.1016/j.antiviral.2013.12.009. Epub 2014 Jan 11. Review. PubMed PMID:24418573. See more on PubMed
Chang MS, Lu YT, Ho ST, Wu CC, Wei TY, Chen CJ, Hsu YT, Chu PC, Chen CH, ChuJM, Jan YL, Hung CC, Fan CC, Yang YC. Antibody detection of SARS-CoV spike andnucleocapsid protein. Biochem Biophys Res Commun. 2004 Feb 20;314(4):931-6.PubMed PMID: 14751221. See more on PubMed
Che XY, Qiu LW, Pan YX, Wen K, Hao W, Zhang LY, Wang YD, Liao ZY, Hua X, ChengVC, Yuen KY. Sensitive and specific monoclonal antibody-based capture enzymeimmunoassay for detection of nucleocapsid antigen in sera from patients withsevere acute respiratory syndrome. J Clin Microbiol. 2004 Jun;42(6):2629-35.PubMed PMID: 15184444; PubMed Central PMCID: PMC427886. See more on PubMed
Cheng M, Chan CW, Cheung RC, Bikkavilli RK, Zhao Q, Au SW, Chan PK, Lee SS,Cheng G, Ho WK, Cheung WT. Cross-reactivity of antibody against SARS-coronavirus nucleocapsid protein with IL-11. Biochem Biophys Res Commun. 2005 Dec23;338(3):1654-60. Epub 2005 Oct 25. PubMed PMID: 16263078. See more on PubMed
Lau SK, Woo PC, Wong BH, Tsoi HW, Woo GK, Poon RW, Chan KH, Wei WI, Peiris JS,Yuen KY. Detection of severe acute respiratory syndrome (SARS) coronavirusnucleocapsid protein in sars patients by enzyme-linked immunosorbent assay. JClin Microbiol. 2004 Jul;42(7):2884-9. PubMed PMID: 15243033; PubMed CentralPMCID: PMC446266. See more on PubMed
Lin Y, Shen X, Yang RF, Li YX, Ji YY, He YY, Shi MD, Lu W, Shi TL, Wang J,Wang HX, Jiang HL, Shen JH, Xie YH, Wang Y, Pei G, Shen BF, Wu JR, Sun B.Identification of an epitope of SARS-coronavirus nucleocapsid protein. Cell Res. 2003 Jun;13(3):141-5. PubMed PMID: 12862314. See more on PubMed
Nakanaga K, Yamanouchi K, Fujiwara K. Protective effect of monoclonalantibodies on lethal mouse hepatitis virus infection in mice. J Virol. 1986Jul;59(1):168-71. PubMed PMID: 3012115; PubMed Central PMCID: PMC253053. See more on PubMed
Rota PA, Oberste MS, Monroe SS, Nix WA, Campagnoli R, Icenogle JP, PeñarandaS, Bankamp B, Maher K, Chen MH, Tong S, Tamin A, Lowe L, Frace M, DeRisi JL, ChenQ, Wang D, Erdman DD, Peret TC, Burns C, Ksiazek TG, Rollin PE, Sanchez A,Liffick S, Holloway B, Limor J, McCaustland K, Olsen-Rasmussen M, Fouchier R,Günther S, Osterhaus AD, Drosten C, Pallansch MA, Anderson LJ, Bellini WJ.Characterization of a novel coronavirus associated with severe acute respiratory syndrome. Science. 2003 May 30;300(5624):1394-9. Epub 2003 May 1. PubMed PMID:12730500. See more on PubMed
Shin GC, Chung YS, Kim IS, Cho HW, Kang C. Preparation and characterization ofa novel monoclonal antibody specific to severe acute respiratorysyndrome-coronavirus nucleocapsid protein. Virus Res. 2006 Dec;122(1-2):109-18.Epub 2006 Aug 30. PubMed PMID: 16942813. See more on PubMed
Surjit M, Liu B, Chow VT, Lal SK. The nucleocapsid protein of severe acuterespiratory syndrome-coronavirus inhibits the activity of cyclin-cyclin-dependentkinase complex and blocks S phase progression in mammalian cells. J Biol Chem.2006 Apr 21;281(16):10669-81. Epub 2006 Jan 23. PubMed PMID: 16431923. See more on PubMed
Surjit M, Liu B, Kumar P, Chow VT, Lal SK. The nucleocapsid protein of theSARS coronavirus is capable of self-association through a C-terminal 209 aminoacid interaction domain. Biochem Biophys Res Commun. 2004 May 14;317(4):1030-6.PubMed PMID: 15094372. See more on PubMed
Tan YJ, Goh PY, Fielding BC, Shen S, Chou CF, Fu JL, Leong HN, Leo YS, Ooi EE,Ling AE, Lim SG, Hong W. Profiles of antibody responses against severe acuterespiratory syndrome coronavirus recombinant proteins and their potential use as diagnostic markers. Clin Diagn Lab Immunol. 2004 Mar;11(2):362-71. PubMed PMID:15013989; PubMed Central PMCID: PMC371215. See more on PubMed
Timani KA, Ye L, Ye L, Zhu Y, Wu Z, Gong Z. Cloning, sequencing, expression,and purification of SARS-associated coronavirus nucleocapsid protein forserodiagnosis of SARS. J Clin Virol. 2004 Aug;30(4):309-12. PubMed PMID:15163419. See more on PubMed
Woo PC, Lau SK, Wong BH, Tsoi HW, Fung AM, Chan KH, Tam VK, Peiris JS, YuenKY. Detection of specific antibodies to severe acute respiratory syndrome (SARS) coronavirus nucleocapsid protein for serodiagnosis of SARS coronavirus pneumonia.J Clin Microbiol. 2004 May;42(5):2306-9. PubMed PMID: 15131220; PubMed CentralPMCID: PMC404667. See more on PubMed

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