Influenza A H7N9 (A/Shanghai/2/2013) Nucleoprotein / NP Insect Cell Lysate (WB positive control)


Influenza A H7N9 (A/Shanghai/2/2013) Nucleoprotein / NP Insect Cell Lysate (WB positive control): 产品信息

This H7N9 Nucleoprotein/NP overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of Nucleoprotein/NP protein (Cat: 40111-V08B) from the overexpression lysate was verified.
Baculovirus-Insect cells
A DNA sequence encoding the Influenza A virus (A/Shanghai/2/2013(H7N9)) nucleoprotein protein (AGL44439.1) (Met1-Asn498) was expressed with a C-terminal polyhistidine tag.
The recombinant nucleoprotein protein of Influenza A virus (A/Shanghai/1/2013(H7N9)) comprises 509 amino acids and has a predicted molecular mass of 57.8 kDa. The apparent molecular mass of the protein is approximately 57 kDa in SDS-PAGE under reducing conditions.

Influenza A H7N9 (A/Shanghai/2/2013) Nucleoprotein / NP Insect Cell Lysate (WB positive control): 使用指南

Cell lysate was prepared by homogenization of the over-expressed cells 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.
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.
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.
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
稳定性 & 储存条件
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.
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.

Influenza A H7N9 (A/Shanghai/2/2013) Nucleoprotein / NP Insect Cell Lysate (WB positive control): 别称

H7N9 NP Overexpression Lysate

Nucleoprotein/NP 背景信息

Influenza A viral nucleoprotein (NP) plays a critical role in virus replication and host adaptation. The influenza A virus nucleoprotein (NP) is an essential multifunctional protein that encapsidates the viral genome and functions as an adapter between the virus and the host cell machinery. NPs from all strains of influenza A viruses contain two nuclear localization signals (NLSs): a well-studied monopartite NLS1 and a less-characterized NLS2, thought to be bipartite. The nucleocapsid is a complex of the viral nucleoprotein, RNA, and several other viral proteins. The nucleoprotein forms large, RNA-bound, helical filaments and acts as a scaffold for additional viral proteins.
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