SARS-CoV-2 (2019-nCoV) Nucleocapsid Insect Cell Lysate (WB positive control)

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SARS-CoV-2 (2019-nCoV) Nucleocapsid Insect Cell Lysate (WB positive control): Product Information

Product Description
This 2019-nCoV Coronavirus Nucleocapsid overexpression lysate was created in Baculovirus-Insect Cells and intented for use as a Western blot (WB) positive control. Purification of Coronavirus Nucleocapsid protein (Cat: 40588-V08B) from the overexpression lysate was verified.
Expression Host
Baculovirus-Insect Cells
Species
2019-nCoV
Sequence Information
A DNA sequence encoding the SARS-CoV-2 (2019-nCoV) Nucleocapsid Protein (YP_009724397.2(335Gly/Ala)) (Met1-Ala419) was expressed with a polyhistidine tag at the C-terminus.
Molecule Mass
The recombinant NCP-CoV(2019-nCoV) Nucleocapsid Protein (His tag) consists of 430 amino acids and predicts a molecular mass of 47.08 kDa.

SARS-CoV-2 (2019-nCoV) Nucleocapsid Insect Cell Lysate (WB positive control): Usage Guide

Preparation Method
Cell lysate was prepared by homogenization in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors (Sigma). Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (Bio-Rad protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
Lysis Buffer
Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.
Recommend Usage
1. Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube. 2. Re-dissolve the pellet using 200μL pure water and boil for 2-5 min. 3. Store the lyophilized cell lysate at 4℃. After re-dissolution, recommend to aliquot it into smaller quantities and store at -80℃.
Sample Buffer
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
Stability & Storage
Store at 4℃ for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80℃ for up to twelve months. Avoid repeated freeze-thaw cycles.
Application
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.

SARS-CoV-2 (2019-nCoV) Nucleocapsid Insect Cell Lysate (WB positive control): Synonyms

2019-nCoV coronavirus NP Overexpression Lysate; 2019-nCoV coronavirus Nucleocapsid Overexpression Lysate; 2019-nCoV coronavirus Nucleoprotein Overexpression Lysate; 2019-nCoV cov np Overexpression Lysate; 2019-nCoV ncov NP Overexpression Lysate; 2019-nCoV NCP-CoV Nucleocapsid Overexpression Lysate; 2019-nCoV novel coronavirus NP Overexpression Lysate; 2019-nCoV novel coronavirus Nucleocapsid Overexpression Lysate; 2019-nCoV novel coronavirus Nucleoprotein Overexpression Lysate; 2019-nCoV np Overexpression Lysate; 2019-nCoV nucleocapsid Overexpression Lysate; 2019-nCoV Nucleoprotein Overexpression Lysate

Coronavirus Nucleocapsid Background Information

Coronaviruses are enveloped viruses with a positive-sense RNA genome and with a nucleocapsid of helical symmetry. Coronavirus nucleoproteins localize to the cytoplasm and the nucleolus, a subnuclear structure, in both virus-infected primary cells and in cells transfected with plasmids that express N protein. Coronavirus N protein is required for coronavirus RNA synthesis, and has RNA chaperone activity that may be involved in template switch. Nucleocapsid protein is a most abundant protein of coronavirus. During virion assembly, N protein binds to viral RNA and leads to formation of the helical nucleocapsid. Nucleocapsid protein is a highly immunogenic phosphoprotein also implicated in viral genome replication and in modulating cell signaling pathways. Because of the conservation of N protein sequence and its strong immunogenicity, the N protein of coronavirus is chosen as a diagnostic tool.
References
    1. 1.Van Boheemen S, et al. (2012), MBio. 3(6):e00473-12.
    2. Bisht H. et al., 2004, Proc Natl Acad Sci. 101 (17): 6641-6.
    3. Li W. et al., 2005, Science. 309 (5742): 1864-8.
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