Influenza A H1N1 (A/Puerto Rico/8/34/Mount Sinai) Nucleoprotein / NP (I116M) Insect Cell Lysate (WB positive control)

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Influenza A H1N1 (A/Puerto Rico/8/34/Mount Sinai) Nucleoprotein / NP (I116M) Insect Cell Lysate (WB positive control): 产品信息

产品描述
This H1N1 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: 11675-V08B) from the overexpression lysate was verified.
表达宿主
Baculovirus-Insect cells
种属
H1N1
蛋白构建信息
A DNA sequence encoding the Influenza A virus (A/Puerto Rico/8/34/Mount Sinai (H1N1)) nucleoprotein (AAM75159.1, with mutation Ile 116 Met) (Met1-Gly490) was fused with a polyhistidine tag at the C-terminus.
分子量
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 56.7 kDa. It migrates as an approximately 50 kDa band in SDS-PAGE under reducing conditions.

Influenza A H1N1 (A/Puerto Rico/8/34/Mount Sinai) Nucleoprotein / NP (I116M) Insect Cell Lysate (WB positive control): 使用指南

制备方法
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.
裂解缓冲液
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.
使用建议
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.
缓冲液
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.
应用
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.

Influenza A H1N1 (A/Puerto Rico/8/34/Mount Sinai) Nucleoprotein / NP (I116M) Insect Cell Lysate (WB positive control): 别称

H1N1 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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