Anti-SARS-CoV-2 nucleocapsid Antibody, clone 5B9-C6

SARS-CoV-2 Nucleoprotein (UniProt: P0DTC9; also known as N, Nucleocapsid protein, NC, Protein N) is encoded by the N gene (Gene ID: 43740575) in SARS-CoV-2 virus. The SARS-CoV-2 is a positive-strand RNA virus that causes severe respiratory syndrome in human. The mature SARS-CoV-2 contains 4 structural proteins: Envelope (E), Membrane (M), Nucleocapsid (N), and the Spike protein (S). E and M proteins help in viral assembly and N protein is needed for RNA synthesis. The S protein is a single-pass type I, homotrimeric, membrane glycoprotein that is responsible for virus binding and entry into host cell. The N protein packages the positive strand viral genome RNA into a helical ribonucleocapsid (RNP) and plays a fundamental role during virion assembly through its interactions with the viral genome and membrane protein M. It plays an important role in enhancing the efficiency of subgenomic viral RNA transcription as well as viral replication. The RNA binding region of SARS-CoV-2 is localized to amino acids 41-186 and its dimerization domain is in amino acids 258-361. Lack of cysteine is one of the common features in nucleoprotein from different coronaviruses. The complete N protein of SARS-CoV and COV-2 is a highly basic with positive net charges and high pI (10.11). Its N-terminal region is more basic with positive charge, while the C-terminal region is acidic with negative charge. The middle region of nucleoprotein displays relatively higher hydrophobicity and the two termini display greater hydrophilicity. Nucleoprotein can undergo phosphorylation at multiple serines and threonines and some of these phosphorylation events are essential for RNA binding, oligomerization and localization to nucleoli. Phosphorylation is also required for recruitment of host RNA helicase DDX1 (DEAD-Box Helicase 1) that facilitates template readthrough and enables longer subgenomic mRNA synthesis. Clone 5B9-C6 is specific for SARS-CoV-2 and does not react with SARS-CoV-1.

Clone 5B9-C6 is a mouse monoclonal antibody that detects nucleocapsid protein of Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). It targets an epitope within the N-terminal domain. It does not cross react with SARS virus.

His-tagged full-length nucleocapsid protein from Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2).

Quality Control Testing

Evaluated by Western Blotting in lysates from HEK293T cells expressing SARS-CoV-2 Nucleocapsid protein.

Western Blotting Analysis (WB): A 1:1,000 dilution of this antibody detected Nucleocapsid protein in lysates from HEK293T cells expressing SARS-CoV-2 Nucleocapsid protein, but not in lysates from the wild type HEK293 T cells.

Tested Applications

Immunoprecipitation Analysis: A representative lot Immunoprecipitated SARS-CoV-2 nucleocapsid protein. (Courtesy of Stefan Schüchner, Ingrid Mudrak, Ingrid Frohner, and Egon Ogris (Max Perutz Labs, Medical University of Vienna, Austria).

ELISA Analysis: A 1:3 dilution from a representative lot detected SARS-CoV-2 nucleocapsid protein. (Courtesy of Stefan Schüchner, Ingrid Mudrak, Ingrid Frohner, and Egon Ogris (Max Perutz Labs, Medical University of Vienna, Austria).

Western Blotting Analysis: A 1:1,000 dilution from a represenative lot detected SARS-CoV-2 nucleocapsid in Whole cell lysates of HEK293T cells expressing the nucleocapsid (NC) proteins of various human coronaviruses (Courtesy of Stefan Schüchner, Ingrid Mudrak, Ingrid Frohner, and Egon Ogris (Max Perutz Labs, Medical University of Vienna, Austria).

Immunofluorescence Analysis: A 1:100 dilution from a represenative lot detected SARS-CoV-2 nucleocapsid in HeLa cells transiently transfected with SARS-CoV-2 NC-P2A-GFP (Courtesy of Stefan Schüchner, Ingrid Mudrak, Ingrid Frohner, and Egon Ogris (Max Perutz Labs, Medical University of Vienna, Austria).

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user

Anti-SARS-CoV-2 nucleocapsid, clone 5B9-C6, Cat. No. MABF2789, is a mouse monoclonal antibody that detects Nucleoprotein in SARS-CoV-2 and is tested for use in ELISA, Immunofluorescence, and Western Blotting.

Purified mouse monoclonal antibody IgG1 in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Recommend storage at +2°C to +8°C. For long term storage antibodies can be kept at -20°C. Avoid repeated freeze-thaws.

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.