Sanne Hoeks

Cytokine assays 89 6 Bio-Rad CST Roche 0 200 400 600 800 1000 Background (MFI) Bio-Rad CST Roche 0 10 20 30 40 50 background/noise ratio * * * * * FIGURE 5. Influence of lysis buffer on assay performance. Peripheral blood human mononuclear cells from healthy donors were isolated using Ficoll-gradient centrifugation. Cells were then stimulated with a concentration 10 ng/ml of LPS for 4hours. Following the incubation period supernatant was removed and cells were washed with PBS. Next cells where lysed according to the lysis protocols for Complete lysis M (Roche), Cell signaling Technologies lysis buffer (CST) and Bio-Rad. The samples where then analyzed by multiplex immunoassay for various markers. The graphs illustrate (left) background levels, only lysis buffer and (right) background - noise ratio for the complete cytokine panel. * indicates all p values <0.05.