Anti-Ebola virus Glycoprotein Magnetic Beads-IP Kit Product Components
Components | Storage |
Anti-Ebola virus Glycoprotein Magnetic Beads1,3 | 2-8℃ for 12 months |
NP40 Cell Lysis Buffer2 | -20℃ for 12 months |
5×TBST(pH7.4) | |
1×TBST(pH7.4) | |
ddH2O | |
CD166 Positive Cell Lysate | -20℃ for 12 months |
Alkaline Elution Buffer | 2-8℃ for 12 months |
Acidity Elution Buffer | 2-8℃ for 12 months |
Neutralization Buffer | 2-8℃ for 12 months |
[1] The IP KIT contains anti-Ebola virus Glycoprotein magnetic Beads (2 mg/mL) in phosphate buffered saline (PBS, pH 7.4) with sodium azide (0.1%).
[2] Using NP-40 cell lysate buffer in the kit is required,otherwise,the magnetic beads may be precipitated.
[3] Shipping: Magnetic Beads kits are shipped at ambient temperature in which magnetic beads are provided in liquid buffer.
Anti-Ebola virus Glycoprotein Magnetic Beads-IP Kit Product Description
The Anti-Ebola virus Glycoprotein magnetic Beads, conjugated with Anti-Ebola virus Glycoprotein antibody, are used for immuneprecipitation (IP) of Ebola virus Glycoprotein proteins which expressed in vitro expression systems. For IP, the beads are added to a sample containing Ebola virus Glycoprotein proteins to form a bead-protein complex. The complex is removed from the solution manually using a magnetic separator. The bound Ebola virus Glycoprotein proteins are dissociated from the magnetic beads using an elution buffer. Anti-Ebola virus Glycoprotein Magnetic Beads-IP Kit Antibody Information
Antibody
Ebola virus EBOV (subtype Zaire, strain Mayinga 1976) Glycoprotein / GP Antibody, Rabbit PAb, Antigen Affinity Purified(
40304-T44)
Immunogen
Recombinant Ebola virus EBOV (subtype Zaire, strain Mayinga 1976) Glycoprotein / GP Protein (Catalog#40304-V08B1)
Species Reactivity
Ebola virus EBOV (subtype Zair
Source
Polyclonal EBOV Rabbit IgG
Preparation
Produced in rabbits immunized with purified, recombinant Ebola virus EBOV (subtype Zaire, strain Mayinga 1976) Glycoprotein / GP (Catalog#40304-V08B1; AAC54887.1; Met1-Gln650), and purified by antigen affinity chromatography.
Applications
Immunoprecipitation (IP), Minimum Protein Purification
Ebola virus Glycoprotein Background Information
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, β-integrins, folate receptor-α, 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.
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.