Recombinant Mouse C8G Protein Purification

Lipopolysaccharide from Escherichia coli 0111:B4 was purchased from Sigma-Aldrich (St. Louis, MO, United States). Recombinant mouse IFN-γ protein was purchased from R&D systems. The bacterially expressed recombinant mouse C8G protein was prepared as described previously (Lee et al., 2008 (link)). Briefly, the recombinant mouse C8G protein without the signal peptide was expressed in E. coli (strain BL21) as a glutathione S-transferase (GST) fusion protein. The protein was purified using glutathione Sepharose 4B beads (GE Healthcare, Chicago, IL, United States). GST was removed by thrombin digestion. C8G was purified and concentrated. SDS-PAGE with Coomassie Brilliant Blue R250 staining showed that the purity of C8G was over 95%. JTE013, S1P, STATTIC, and BAY 11-7082 were purchased from Cayman Chemical (Ann Arbor, MI, United States). CYM-5478 was purchased from Aobious Inc. (Gloucester, MA, United States).