SARS-CoV
SARS-CoV Nucleocapsid (422a.a) Recombinant

Introduction:
SARS Coronavirus is an enveloped virus containing three outer structural proteins, namely the membrane (M), envelope (E), and spike (S) proteins. Spike (S)-glycoprotein of the virus interacts with a cellular receptor and mediates membrane fusion to allow viral entry into susceptible target cells. Accordingly, S-protein plays an important role in virus infection cycle and is the primary target of neutralizing antibodies.

Description:
The Recombinant SARS-CoV Nucleocapsid Protein is manufactured with N-terminal fusion HisTag. The Recombinant SARS-CoV Nucleocapsid His-Tagged Fusion Proteinis 47.8 kDa containing 422 amino acid residues of the SARS-CoV Nucleocapsid protein and 15 additional amino acid residues – HisTag (underlined).
MRGSHHHHHH GMASHMSDNG PQSNQRSAPR ITFGGPTDST DNNQNGGRNG ARPKQRRPQG LPNNTASWFT ALTQHGKEEL RFPRGQGVPI NTNSGPDDQI GYYRRATRRV RGGDGKMKEL SPRWYFYYLG TGPEASLPYG ANKEGIVWVA TEGALNTPKD HIGTRNPNNN AATVLQLPQG TTLPKGFYAE GSRGGSQASS RSSSRSRGNS RNSTPGSSRG NSPARMASGG GETALALLLL DRLNQLESKV SGKGQQQQGQ TVTKKSAAEA SKKPRQKRTA TKQYNVTQAF GRRGPEQTQG NFGDQDLIRQ GTDYKHWPQI AQFAPSASAF FGMSRIGMEV TPSGTWLTYH GAIKLDDKDP QFKDNVILLN KHIDAYKTFP PTEPKKDKKK KTDEAQPLPQ RQKKQPTVTL LPAADMDDFS RQLQNSMSGA SADSTQA

Specificity:
The amino acid sequence of the recombinant SARS-CoV Nucleocapsid protein is 100% homologous to amino acid sequence of the native SARS-CoV Nucleocapsid protein.

Purification Method:
Three-step procedure using affinity Ni-NTA chromatography and size exclusion chromatography before and after refolding

Source:
E.coli

Formulation:
Sterile filtered and lyophilized from 0.5 mg/ml in 0.05 M Acetate buffer pH4 Reconstitution: Add 0.2 ml of 0.1M Acetate buffer pH4 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.

Purity:
>95% (SDS-PAGE analyzed).

Storage:
Store lyophilized protein at -20°C. Aliquot the product after reconstitution to avoid repeated freezing/thawing cycles. Reconstituted protein can be stored at 4°C for a limited period of time; it does not show any change after two weeks at 4°C.

Stability/Shelf Life:
The lyophilized protein remains stable until the expiry date when stored at -20°C.

Applications:
Western blotting

Usage:
ProSpec's products are furnished for LABORATORY RESEARCH USE ONLY. The product may not be used as drugs, agricultural or pesticidal products, food additives or household chemicals.

References:
• Che XY, Qiu LW, Pan YX, Wen K, Hao W, Zhang LY, Wang YD, Liao ZY, Hua X, Cheng VC, Yuen KY. Sensitive and specific monoclonal antibody-based capture enzyme immunoassay for detection of nucleocapsid antigen in sera from patients with severe acute respiratory syndrome. J Clin Microbiol. 2004 Jun;42(6):2629-35.

• 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 coronaviral nucleocapsid or spike proteins by immunoblotting or ELISA. J Med Virol. 2004 Jul;73(3):338-46.

• Timani KA, Ye L, Ye L, Zhu Y, Wu Z, Gong Z. Cloning, sequencing, expression, and purification of SARS-associated coronavirus nucleocapsid protein for serodiagnosis of SARS. J Clin Virol. 2004 Aug;30(4):309-12.

• Woo PC, Lau SK, Wong BH, Tsoi HW, Fung AM, Chan KH, Tam VK, Peiris JS, Yuen KY. 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.

• 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. 2004 Apr 2;316(2):476-83.

• 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 acute respiratory syndrome coronavirus
recombinant proteins and their potential use as diagnostic markers. Clin Diagn Lab Immunol. 2004 Mar;11(2):362-71.

• Chang MS, Lu YT, Ho ST, Wu CC, Wei TY, Chen CJ, Hsu YT, Chu PC, Chen CH, Chu JM, Jan YL, Hung CC, Fan CC, Yang YC. Antibody detection of SARS-CoV spike and nucleocapsid protein. Biochem Biophys Res Commun. 2004 Feb 20;314(4):931-6.

• 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 SARScoronavirus nucleocapsid protein. Cell Res. 2003 Jun;13(3):141-5.
. Cell Res. 2003 Jun;13(3):141-5.