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

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Vaccinia Virus B18R Transfected / Overexpression Cell Lysate Product Information
Expressed Host:Baculovirus-Insect cells
Product Description:Baculovirus-Insect Cell lysate that Vaccinia Virus B18R transfected / overexpressed for Western blot (WB) positive control. The whole cell lysate is provided in 1X Sample Buffer (1X modified RIPA buffer+1X SDS loading buffer).
Sequence information:A DNA sequence encoding the B18R (P25213) (Met1-Glu351) was expressed with a C-terminal polyhistidine tag.
Predicted N Terminal:His 20
Molecule Mass: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.
Species:Vaccinia Virus
Vaccinia Virus B18R Transfected / Overexpression Cell Lysate Usage Guide
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 (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.
Lysis 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.
Quality Control Testing:12.5% SDS-PAGE Stained with Coomassie Blue after protein purification.
Stability:Samples are stable for up to twelve months from date of receipt.
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 200μL pure water and boil for 2-5 min. 3. Store the lyophilized cell lysate at 4℃. After re-dissolution, recommend to aliquot it into smaller quantities and store at -80℃.
Storage Buffer:1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
Storage Instruction:Store at 4℃. After re-dissolution, aliquot and store at -80℃.
Application notes:Western blot (WB): Use at an assay dependent dilution.
Other Applications: Not tested.
Optimal dilutions/concentrations should be determined by the end user.
B18R / B19R Background

B18R is a type I interferon (IFN)-binding protein, which is encoded by the B18R open reading frame in the WR (Western Reserve)strain of vaccinia virus. It is also known as B19R in the Copenhagen strain of Vaccinia. 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.

Vaccinia Virus B18R / B19R References
  1. I Vancov. et al., 1998, J Gen Virol July. 79 (7): 1647-9.
  2. Colamonici OR. et al., 1995, J Biol Chem. 270 (27): 15974-8.
  3. Goebel SJ. et al., 1990, Virology 179: 247-66.
  4. Goebel SJ. et al., 2010, J Cell Biol. 190 (4): 533-9.
Size / Price
Catalog: 40020-V08BL-300
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