Journal:

Article Title: Acute-Phase Concentrations of Lipopolysaccharide (LPS)-Binding Protein Inhibit Innate Immune Cell Activation by Different LPS Chemotypes via Different Mechanisms

doi: 10.1128/IAI.73.1.193-200.2005

Figure Lengend Snippet: hd-LBP inhibits binding of R and S LPS chemotypes to sCD14. Binding of R-type LPS from E. coli F515 (Re-LPS) (A), R-type LPS from S. enterica serovar Typhimurium TV 119 (Ra-LPS) (B), and S-type LPS from E. coli O111:B4 (C) to recombinant human sCD14 and the modulating effects of increasing concentrations of LBP were analyzed by native PhastGel electrophoresis. Following incubation at 37°C for 10 min, samples were subjected to automated native electrophoresis on 20% PhastGels and silver staining by using the PhastSystem apparatus. The positions of uncomplexed sCD14, sCD14-LPS complexes, LBP, and the LPS preparations are indicated. Arrows indicate different sCD14-LPS complexes containing most likely LPS of different chain lengths (C). As a loading control, the native samples were subsequently adjusted to a final concentration of 2% (wt/vol) SDS and additionally analyzed by PhastGel SDS electrophoresis and silver staining (lower panels).

Article Snippet: Briefly, R-type LPS from E. coli F515 or S-type LPS from E. coli O111:B4 at a concentration of 0.025 or 0.113 μg/μl, respectively, was incubated in Dulbecco's phosphate-buffered saline (without magnesium or calcium; Life Technologies/GIBCO BRL, Eggenstein, Germany) in the absence or presence of 0.25 μg of recombinant human sCD14 (Biometec, Greifswald, Germany)/μl at 37°C for 10 min. For analysis of LBP effects, recombinant human LBP (Xoma Corp.) was added at final concentrations of 0.0005, 0.0025, 0.025, 0.125, and 0.25 μg/μl.

Techniques: Binding Assay, Recombinant, Electrophoresis, Incubation, Silver Staining, Control, Concentration Assay