Zika virus (ZIKV) (strain Zika SPH2015) ZIKV-NS5 Insect Cells Cell Lysate (WB positive control)

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Zika virus (ZIKV) (strain Zika SPH2015) ZIKV-NS5 Insect Cells Cell Lysate (WB positive control): Product Information

Product Description
This ZIKV ZIKV Nonstructural protein 5 overexpression lysate was created in Baculovirus-Insect Cells and intented for use as a Western blot (WB) positive control. Purification of ZIKV Nonstructural protein 5 protein (Cat: 40546-V08B) from the overexpression lysate was verified.
Expression Host
Baculovirus-Insect Cells
Species
ZIKV
Sequence Information
A DNA sequence encoding the Zika virus (ZIKV) (strain ZikaSPH2015) NS5 (ALU33341.1) (Lys2772-Leu3423) was expressed with a polyhistidine tag at the C-terminus.
Molecule Mass
The recombinant Zika virus (ZIKV) (strain ZikaSPH2015) NS5 consists of 663 amino acids and predicts a molecular mass of 76.7 kDa.

Zika virus (ZIKV) (strain Zika SPH2015) ZIKV-NS5 Insect Cells 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.

ZIKV Nonstructural protein 5 Background Information

Zika virus NS5 is invloved in methytransferase and RNA guanylytransferase activities and capping and synthesis of RNA. And, NS5 is also a RNA-dependent RNA polymerase.
References
    1. Wong SS-Y, et al., Zika virus infectiondthe next wave after dengue?, Journal of the Formosan Medical Association (2016)
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