Human respiratory syncytial virus (RSV) (A, rsb1734) glycoprotein G / RSV-G HEK293 Cell Lysate (WB positive control): 产品信息
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 (Cat: 11070-V08H) from the overexpression lysate was verified.
A DNA sequence encoding the glycoprotein G extracellular domain (Asn 66-Arg 297) of human respiratory syncytial virus A (95 % homologous with strain rsb1734) (P27022-1) was expressed, with a C-terminal polyhistidine tag.
The secreted recombinant human RSV-G comprises 242 amino acids with a predicted molecular mass of 26.7 kDa. As a result of highly glycosylation, the RSV-G protein migrates as an approximately 80-90 kDa band in SDS-PAGE under reducing conditions.
Human respiratory syncytial virus (RSV) (A, rsb1734) glycoprotein G / RSV-G HEK293 Cell Lysate (WB positive control): 使用指南
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.
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.
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
稳定性 & 储存条件
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.
Western Blot (WB) Optimal dilutions/concentrations should be determined by the end user.
Human respiratory syncytial virus (RSV) (A, rsb1734) glycoprotein G / RSV-G HEK293 Cell Lysate (WB positive control): 别称
RSV G Overexpression Lysate
RSV Glycoprotein G 背景信息
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 8-9 kDa. 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.
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