respiratory syncytial virus (RSV) Lysate | RSV-G lysate

Human respiratory syncytial virus (RSV)(subtype A, strain Long) Glycoprotein G HEK293 Cell Lysate (WB positive control)

• Applications: Western Blot
• Synonyms: respiratory syncytial virus (RSV); Human respiratory syncytial virus (RSV)(subtype A; strain Long) Glycoprotein G HEK293 Cell Lysate (WB positive control); RSV G Overexpression Lysate; RSV-G lysate

• Host: HEK293 Cells
• Form/Format: Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.; 1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).

• Applicable Applications for RSV-G lysate: Western Blot (WB)
• Species: RSV
• Sequence Information: A DNA sequence encoding the human respiratory syncytial virus (RSV)(subtype A, strain Long) glycoprotein G (P20895-1)(His67-Gln298) was expressed with a polyhistidine tag at the C-terminus.
• 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 (protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50mM Tris-HCl pH6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
• 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.; 3. Store the lyophilized cell lysate at 4 degree C. After re-dissolution, recommend to aliquot it into smaller quantities and store at -80 degree C.
• 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.

Related Product Information for RSV-G lysate

Description: This RSV RSV Glycoprotein G overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of RSV Glycoprotein G protein from the overexpression lysate was verified.

Background: Human respiratory syncytial virus (HRSV) is the most common etiological agent of acute lower respiratory tract disease in infants and can cause repeated infections throughout life. It is classified within the genus pneumovirus of the family paramyxoviridae. Like other members of the family, HRSV has two major surface glycoproteins (G and F) that play important roles in the initial stages of the infectious cycle. HRSV G protein is a type II glycoprotein of 289-299 amino acids (depending on the virus strain) with a signal/anchor hydrophobic domain and is extensively modified by the addition of both N-and O-linked oligosaccharides to achieve the mature form of 8-9kDa. The C-terminal ectodomain of the G protein has a central region and four cysteines which are conserved in all HRSV isolates and have been proposed as the putative receptor binding site. The G protein mediates attachment of the virus to the host cell membrane by interacting with heparan sulfate, initiating the infection. As similar to mucins in amino acid compositions, the RSV G protein can interact with host CX3CR1, the receptor for the CX3C chemokine fractalkine, and thus modulates the immune response and facilitate infection. Secreted glycoprotein G helps RSV escape antibody-dependent restriction of replication by acting as an antigen decoy and by modulating the activity of leukocytes bearing Fcgamma receptors. Unlike the other paramyxovirus attachment proteins, HRSV-G lacks both neuraminidase and hemagglutinating activities.

References

Martin-Gallardo A. et al., 1993, J Gen Virol. 74 : 453-8. Jose AM. et al.,1997, J Gen Virol. 78: 2411-8. Feldman SA. et al., 1999, J Virol. 73: 6610-7. Garca-a-Beato R. et al., 2000, J Gen Virol. 81: 919-27. Zlateva KT. et al., 2004, J Virol. 78: 4675-83. Trento A. et al., 2006, J Virol. 80: 975-84.