Zika virus (ZIKV) (strain Zika SPH2015) E / Envelope protein (aa 291-696, His Tag) Insect Cell Lysate (WB positive control)

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Zika virus (ZIKV) (strain Zika SPH2015) E / Envelope protein (aa 291-696, His Tag) Insect Cell Lysate (WB positive control): Product Information

Product Description
This ZIKV ZIKV Envelope overexpression lysate was created in Baculovirus-Insect Cells and intented for use as a Western blot (WB) positive control. Purification of ZIKV Envelope protein (Cat: 40543-V08B5) from the overexpression lysate was verified.
Expression Host
Baculovirus-Insect Cells
Species
ZIKV
Sequence Information
A DNA sequence encoding the Zika virus (strain Zika SPH2015) E (ALU33341.1, with mutation Thr 356 Ala, Gln 357 Gly, Trp 381 Arg, Leu 387 Arg) (Ile291-Thr696) was expressed with a polyhistidine tag at the C-terminus.
Molecule Mass
The recombinant Zika virus (strain Zika SPH2015) E consists of 417 amino acids and predicts a molecular mass of 45.6 kDa.

Zika virus (ZIKV) (strain Zika SPH2015) E / Envelope protein (aa 291-696, His Tag) 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.

ZIKV Envelope Background Information

Envelope of Zika virus is resposible for receptor binding and membrane. Analysis of the envelope protein of Zika, from Brazilian Zika SPH215 (KU321639), indicates predicted B and T cell epitopes in peptides that are consistent to those reported for dengue, YFYF and Japanese encephalitis. The envelope Domain II B cell epitope, to which much dengue non-neutralizing cross reaction is attributed, is also conserved also in Zika virus, consistent with prior field observations of cross reactivity with dengue and YF. Domain III of the Zika envelope protein, likely the main specific neutralizing domain, is distinct from recent Brazilian dengue isolates and a recent Peruvian YF isolate (GQ379163), 76% of possible major histocompatibility complex class (MHC) I and MHC II binding peptides and potential B cell linear epitopes are unique to Zika virus.
References
  • Marina Basarab specialty registrar. et al .Zika virus. BMJ 2016; 352:i1049 doi: 10.1136/bmj.i1049
  • Pinto Júnior VL, et al. Zika virus: a review to clinicians, Acta Med Port 2015 Nov-Dec;28(6):760-765
  • L Zammarchi, et al.Zika virus infection in a traveler returning to Europe from Brazil, March 2015
  • Arturo Galindo-Fraga, et al. Zika virus: A new epidemic on our doorstep. Rev Inves Clin.2015, 67: 329-32
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