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. 2001 Jan 16;98(2):603–608. doi: 10.1073/pnas.98.2.603

Figure 7.

Figure 7

(a) Specific activity of rBoCD14 depends on method of isolation. The indicated concentrations of rBoCD14 affinity purified by using mAbs 3C10 or 12CA5, or rabbit anti-nBoCD14 was used to stimulate Igκ expression by 70Z/3 as described in Materials and Methods. (Inset) An immunoblot of sCD14 isolated on each of the affinity columns revealed by using mAb 12CA5 followed by HRP-conjugated goat-anti-mouse IgG. (b) PMB inhibits LPS-mediated costimulation of low specific activity rBoCD14. rBoCD14 (0.3 μg/ml) isolated on 12CA5 was preincubated with 100 ng/ml LPS at 37°C for 1 h in serum-free medium followed by addition of 30 μg/ml PMB. After 1 h of incubation at 37°C, 70Z/3 cells were added, and Igκ expression was assessed 22 h later as described in Materials and Methods.