Cytotoxicity Bioassay Analysis of Murine TNF-alpha was performed by resuspending 929 cells in 10% Horse Serum. Six thousand cells per well (passage 8, in 100 µL volume) were transferred to assay plate and incubated overnight at 37°C. Mouse TNF-alpha Recombinant Protein (Product # RMTNFAI), was serially diluted 1:4 in triplicate starting from a concentration of 100 ng/mL in assay media containing 1 µg/mL Actinomycin D, 2X Gentamicin and 10% Horse Serum, and 100 µL was added to each well. The plate was incubated for 23 hours at 37°C and then analyzed via a colorimetric method to determine cell viability and read at 490 nm.