Recombinant IL-1a mouse was spiked into 50% mouse serum and 50% mouse plasma and diluted in a 2-fold dilution series in Sample Diluent NS. The concentrations of mouse IL-1a were measured in duplicate and interpolated from the mouse IL-1a standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are graphed (mean +/- SD).

RAW264.7 cells were cultured in high glucose DMEM with 10% fetal calf serum or horse serum, 2 mM L-glutamine and 100 µg/mL Kanamycin. RAW264.7 cells were starved for 24 hours and treated in the presence and absence of 5 µg/mL of LPS. The concentrations of IL1-a were interpolated from the calibration curve and corrected for sample dilution (1:2). The mean IL-1a concentration in RAW264.7 media was determined to be 288 pg/mL in stimulated RAW 264.7 cells and undetectable in unstimulated RAW264.7 cells.