Ultra low endotoxin fbs

Mini-PROTEAN^® TGX™ precast gels 4–20%, Precision Plus Protein™ Unstained Standards, Clarity™ Western ECL Substrate, and DC™ protein assay were bought from Bio-Rad Laboratories (Segrate, Milano, Italy). Ultra-low Endotoxin FBS was purchased from Euroclone (Pero, Milano, Italy). Rabbit anti-TLR4 polyclonal FITC-conjugated antibody was purchased from StressMarq. Rabbit monoclonal anti-NF-kB (#8242), rabbit monoclonal anti-p-NF-kB (#3031), rabbit monoclonal anti-NLRP3 (#15101), rabbit monoclonal anti-iNOS (#13120) or anti-rabbit IgG, HRP-linked (#7074) antibodies were purchased from Cell Signalling Technology (Danvers, MA, USA). ELISA kit for TNF-α quantification and Griess (G4410) reagent for nitrite quantification was from Sigma-Aldrich (Saint Louis, MO, USA).
Deuterium oxide (D₂O, 99.90% D) and CD₃OD (99.80% D) were purchased from Eurisotop (Cambridge Isotope Laboratories, Inc., Saint-Aubin, France). Lipopolysaccharide (LPS) from E. coli O127:B8, mouse anti-tubulin antibody, FITC-conjugated LPS from E. coli O111:B4, standard 3-(trimethylsilyl)-propionic-2,2,3,3-d4 acid sodium salt (TMSP), sodium phosphate dibasic anhydrous, sodium phosphate monobasic anhydrous, phosphate-buffered saline (PBS), Dulbecco’s phosphate-buffered saline (DPBS), Accutase and all other solvents and chemicals were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA).

Deuterium oxide (H2O-d2, 99.90% D) and MeOH-d4 (99.80% D) were purchased from Eurisotop (Cambridge Isotope Laboratories, Inc, Saint-Aubin, France). Ultra-low Endotoxin FBS was obtained from Euroclone (Euroclone, Milan, Italy). Mini-PROTEAN^® TGX™ precast gels 4–20%, Precision Plus Protein™ Unstained Standards, Clarity™ Western ECL Substrate and DC™ protein assay were purchased from Bio-Rad Laboratories (Hercules, California, United States). Primary antibodies against Nf-kB were purchased from Millipore (Merck Millipore, Burlington, Massachusetts, United States). Standard 3-(trimethylsilyl)-propionic-2,2,3,3-d4 acid sodium salt (TMSP), sodium phosphate dibasic anhydrous sodium phosphate monobasic anhydrous, Dulbecco’s modified Eagle medium (DMEM), penicillin, streptomycin, glutamine, lipopolysaccharide (LPS) from Escherichia coli serotype O127:B8, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), and all the other solvents and chemicals were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA).

BV-2 murine microglial cells were kindly provided by Prof. Elisabetta Blasi (University of Modena and Reggio Emilia, Modena, Italy) and were cultured in DMEM supplemented with 10% heat inactivated Ultra-low Endotoxin FBS (Euroclone, Milano, Italy), L-glutamine (1%), and streptomycin (1%) in a humidified incubator maintained at 37 °C and 5% CO₂, according to Blasi et al. [41 (link)].