EBOV EBOV-G/Glycoprotein Cell Lysate | EBOV-G cell lysate
Ebola virus EBOV (subtype Zaire, strain H.sapiens-wt/SLE/2014/ManoRiver-G3686.1) EBOV-G/Glycoprotein HEK293 Cell Lysate (WB positive control)
Applications
Western Blot
Synonyms
EBOV EBOV-G/Glycoprotein; Ebola virus EBOV (subtype Zaire; strain H.sapiens-wt/SLE/2014/ManoRiver-G3686.1) EBOV-G/Glycoprotein HEK293 Cell Lysate (WB positive control); EBOV-G cell lysate
Host
HEK293 Cells
Form/Format
1x Sample Buffer (1x modified RIPA buffer+1x SDS loading 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.
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.
Sequence Positions
629aa
Sequence
Met1-Gln650, Ser46Asn, Phe291Arg
Applicable Applications for EBOV-G cell lysate
Western Blot (WB)
Application Notes
WB: Use at an assay dependent dilution.
Species
Ebola virus
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 200uL pure water and boil for 2-5 min.
Sequence Construction
A DNA sequence encoding the Zaire ebolavirus(strain Ebola virus H.sapiens-wt/SLE/2014/ManoRiver-G3686.1) GP (AIE11917.1) (Met1-Gln650, Ser46Asn, Phe291Arg) was expressed with a polyhistidine tag at the C-terminus.
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. Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
Preparation and Storage
Store at 4 degree C for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80 degree C for up to twelve months. Avoid repeated freeze-thaw cycles.
Samples are stable for up to twelve months from date of receipt.
Samples are stable for up to twelve months from date of receipt.
Related Product Information for EBOV-G cell lysate
The fourth gene of the EBOV genome encodes a 16-kDa envelope-attached glycoprotein (GP) and a 11 kDa secreted glycoprotein (sGP). Both GP and sGP have an identical 295-residue N-terminus, however, they have different C-terminal sequences. Recently, great attention has been paid to GP for vaccines design and entry inhibitors isolation. GP is a class I fusion protein which assembles as trimers on viral surface and plays an important role in virus entry and attachment. Mature GP is a disulfide-linked heterodimer formed by two subunits, GP1 and GP2, which are generated from the proteolytical process of GP precursor (pre-GP) by cellular furin during virus assembly. The GP1 subunit contains a mucin domain and a receptor-binding domain (RBD); the GP2 subunit has a fusion peptide, a helical heptad-repeat (HR) region, a transmembrane (TM) domain, and a 4-residue cytoplasmic tail. The RBD of GP1 mediates the interaction of EBOV with cellular receptor (e.g. DC-SIGN/LSIGN, TIM-1, hMGL, NPC1, beta-integrins, folate receptor-alpha, and Tyro3 family receptors), of which TIM1 and NPC1 are essential for EBOV entry; the mucin domain having N-and O-linked glycans enhances the viral attachment to cellular hMGL, and participates in shielding key neutralization epitopes, which helps the virus evades immune elimination. There are large conformation changes of GP2 during membrane fusion, which enhance the insertion of fusion loop into cellular membrane and facilitate the release of viral nucleocapsid core to cytoplasm.
This EBOV Ebola virus Glycoprotein overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Ebola virus Glycoprotein protein from the overexpression lysate was verified.
This EBOV Ebola virus Glycoprotein overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Ebola virus Glycoprotein protein from the overexpression lysate was verified.
References
Volchkov VE, et al. Processing of the Ebola virus glycoprotein by the proprotein convertase furin. Proc Natl Acad Sci U S A. 1998 May 12;95(10):5762-7. Lee JE, et al. Structure of the Ebola virus glycoprotein bound to an antibody from a human survivor. Nature. 2008 Jul 10;454(7201):177-82. doi: 10.1038/nature07082. Hood CL, et al. Biochemical and structural characterization of cathepsin L-processed Ebola virus glycoprotein: implications for viral entry and immunogenicity. J Virol. 2010 Mar;84(6):2972-82. doi: 10.1128/JVI.02151-09. Cook JD and Lee JE. The secret life of viral entry glycoproteins: moonlighting in immune evasion. PLoS Pathog. 2013 May;9(5):e1003258. doi: 10.1371/journal.ppat.1003258. Miller EH and Chandran K. Filovirus entry into cells-new insights. Curr Opin Virol. 2012 Apr;2(2):206-14. doi: 10.1016/j.coviro.2012.02.015.
NCBI and Uniprot Product Information
Molecular Weight
The recombinant Zaire ebolavirus (strain Ebola virus H.sapiens-wt/SLE/2014/ManoRiver-G3686.1) consists 629 amino acids and predicts a molecular mass of 69.3kDa.
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Product Notes
The EBOV-G (Catalog #AAA8116060) is a Cell Lysate produced from HEK293 Cells and is intended for research purposes only. The product is available for immediate purchase. The immunogen sequence is 629aa. AAA Biotech's EBOV EBOV-G/Glycoprotein can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB). WB: Use at an assay dependent dilution. Researchers should empirically determine the suitability of the EBOV-G for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. The amino acid sequence is listed below: Met1-Gln65 0, Ser46Asn, Phe291Arg. It is sometimes possible for the material contained within the vial of "EBOV EBOV-G/Glycoprotein, Cell Lysate" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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