Recombinant Viral B8R Biotinylated Protein, CF Summary
Lys18-Ser272, with a C-terminal 6-His tag
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
|Formulation||Lyophilized from a 0.2 μm filtered solution in PBS.|
|Reconstitution||Reconstitute at 250 μg/mL in PBS.|
|Shipping||The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.|
|Stability & Storage:||Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
Both Biotinylated Recombinant Viral B8R (Catalog # BT8225) and unlabeled Recombinant Viral B8R (Catalog # 8225-BR) inhibits IFN-gamma-mediated protection of HeLa human cervical epithelial carcinoma cells to viral lysis. The ED50 for this effect is 0.5-3 ng/mL. The similarity in activity highlights that the biotinylated protein is fully functional.
B8R is a secreted Interferon gamma (IFN-gamma ) binding protein encoded by the open reading frame of the Vaccina virus (smallpox) (1, 2). B8R is a 43 kDa protein and shares only 19% amino acid sequence identity with the extracellular region of the human IFN-gamma Receptor 1 (IFN-gamma R1). However, B8R binds IFN-gamma orthologs from multiple species including human, rat, rabbit, bovine, and chicken with high affinity, but it binds mouse IFN-gamma with low affinity (1, 3, 4). During infection,
virally-induced B8R binds and sequesters endogenous IFN-gamma, thereby suppressing the host immune response and promoting viral immune evasion (5). While B8R is not required for viral replication, it contributes significantly to Vaccinia virus virulence and is frequently inactivated prior to Vaccinia use in vaccines (5-9). IFN-gamma peptide mimetics that activate IFN-gamma R1 but are not bound and sequestered by B8R have been developed for research and therapeutic use (10, 11).
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- Smith, G.L. et al. (2013) J. Gen. Virol. 94:2367.
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- Denes, B. et al. (2006) J. Gene Med. 8:814.
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- Ahmed, C.M. et al. (2007) J. Immunol. 178:4576.
- Ahmed, C.M. et al. (2005) J. Virol. 79:5632.
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