Vaccinia Virus B18R / B19R Insect Cell Lysate (WB positive control)

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Vaccinia Virus B18R / B19R Insect Cell Lysate (WB positive control): 产品信息

产品描述
This Vaccinia Virus Vaccinia virus B18R overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of Vaccinia virus B18R protein (Cat: 40020-V08B) from the overexpression lysate was verified.
表达宿主
Baculovirus-Insect cells
种属
Vaccinia Virus
蛋白构建信息
A DNA sequence encoding the B18R (P25213) (Met1-Glu351) was expressed with a C-terminal polyhistidine tag.
分子量
The secreted recombinant B18R consists of 342 amino acids and predicts a molecular mass of 39.8 KDa. The apparent molecular mass of the protein is approximately 46 KDa in SDS-PAGE under reducing conditions due to glycosylation.

Vaccinia Virus B18R / B19R Insect Cell Lysate (WB positive control): 使用指南

制备方法
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.
裂解缓冲液
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.
使用建议
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.
缓冲液
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.
应用
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.

Vaccinia virus B18R 背景信息

B18R is a type I interferon (IFN)-binding protein, which is encoded by the B18R open reading frame in the WR (WesternReserve)strain of vaccinia virus. Itis alsoknownasB19RintheCopenhagenstrainofVaccinia. B18R exists in a soluble and a membrane-bound form. As a type I IFN receptor, B18R has a broad species specificity. It has high affinity for human IFN-alpha and also binds rabbit, bovine, rat, pig, and mouse IFN-alpha and IFN-beta. It has been shown that secreted B18R binds to uninfected and infected cells. It presents at the cell surface and protects cells from the antiviral state induced by IFN-alpha and IFN-beta. Binding of soluble recombinant B18R protects cultured cells from IFN and allows vaccinia virus replication.
全称
bone marrow stromal cell antigen 2
参考文献
  • I Vancov. et al., 1998, J Gen Virol July. 79 (7): 1647-9.
  • Colamonici OR. et al., 1995, J Biol Chem. 270 (27): 15974-8.
  • Goebel SJ. et al., 1990, Virology 179: 247-66.
  • Goebel SJ. et al., 2010, J Cell Biol. 190 (4): 533-9.
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