Influenza A H1N1 Nucleoprotein / NP Insect Cell Lysate (WB positive control)

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Influenza A H1N1 Nucleoprotein / NP Insect Cell Lysate (WB positive control): Product Information

Product Description
This H1N1 Nucleocapsid / NP overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of Nucleocapsid / NP protein (Cat: 11675-V08B) from the overexpression lysate was verified.
Expression Host
Baculovirus-Insect cells
Species
H1N1
Sequence Information
A DNA sequence encoding the Influenza A virus (A/Puerto Rico/8/34/Mount Sinai (H1N1)) nucleoprotein (AAM75159.1) (Met1-Gly490) was fused with a polyhistidine tag at the C-terminus.
Molecule Mass
The recombinant Influenza A virus (A/Puerto Rico/8/34/Mount Sinai (H1N1)) nucleoprotein comprises 501 amino acids and has a predicted molecular mass of 56.6 kDa. It migrates as an approximately 50 kDa band in SDS-PAGE under reducing conditions.

Influenza A H1N1 Nucleoprotein / NP Insect Cell Lysate (WB positive control): Usage Guide

Preparation Method
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.
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.
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.

Influenza A H1N1 Nucleoprotein / NP Insect Cell Lysate (WB positive control): Alternative Names

H1N1 NP Overexpression Lysate

Nucleocapsid / NP Background Information

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.
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