Journal: Journal of Lipid Research

Article Title: Assessment of cholesteryl ester transfer protein inhibitors for interaction with proteins involved in the immune response to infection [S]

doi: 10.1194/jlr.M002295

Figure Lengend Snippet: Effects of CETP on LPS binding to LBP and BPI. Purified CETP was added to wells containing immobilized LBP or BPI followed by addition of biotinylated LPS to achieve the final CETP concentrations indicated. Results for LPS binding to LBP (A) and BPI (B) represent the mean for four experiments each. * For effects on LPS-LBP binding at 10 nM CETP relative to control P = 0.045.

Article Snippet: Purified recombinant human LBP was purchased from R and D Systems.

Techniques: Binding Assay, Purification, Control

Journal:

Article Title: Basis for the Failure of Francisella tularensis Lipopolysaccharide To Prime Human Polymorphonuclear Leukocytes

doi: 10.1128/IAI.02011-05

Figure Lengend Snippet: F. tularensis LVS LPS competed poorly with N. meningitidis LOS for binding to adsorbed LBP. Recombinant human LBP was adsorbed to 96-well plates, and N. meningitidis (Nm) [3H]LOS (5 ng/ml) was incubated in the wells for 3 hours at 37°C in the presence of 0.1% BSA. Unlabeled N. meningitidis LOS or F. tularensis (Ft) LVS LPS was coincubated at the indicated excess concentrations. Binding of N. meningitidis [3H]LOS was measured as counts per minute in duplicates and was normalized within each experiment. Counts per minute in the absence of unlabeled LPS (or LOS) were defined as 100% (i.e., the control value). Background counts per minute were subtracted from the results for each sample. P < 0.001 for comparisons between F. tularensis LVS LPS and N. meningitidis LOS at 10- and 100-fold excess concentrations. Error bars are the standard errors of the means; n = 3.

Article Snippet: Rabbit antiserum to recombinant human LBP (Synpep, Dublin, CA) was purified for immunoglobulins with a HiTrap rProtein A affinity column as per the manufacturer's instructions (Amersham Biosciences).

Techniques: Binding Assay, Recombinant, Incubation, Control

Journal:

Article Title: Basis for the Failure of Francisella tularensis Lipopolysaccharide To Prime Human Polymorphonuclear Leukocytes

doi: 10.1128/IAI.02011-05

Figure Lengend Snippet: F. tularensis LVS LPS did not compete for binding to LBP in solution. N. meningitidis (Nm) [3H]LOS (3 ng/ml) was coincubated with unlabeled N. meningitidis LOS, F. tularensis (Ft) LVS LPS, msbB LOS, or buffer for 30 min at 37°C with a limiting concentration of LBP (1 ng/ml) and 0.1% BSA. The resulting N. meningitidis [3H]LOS-LBP complexes were then incubated in duplicate in wells coated with polyclonal rabbit anti-human LBP IgG or preimmune IgG. Binding of N. meningitidis [3H]LOS was measured as counts per minute in duplicates and was normalized within each experiment after subtraction of counts from preimmune antibody-coated wells. Counts per minute in the absence of unlabeled LPS (or LOS) were defined as 100% (i.e., the control value) P < 0.001 for comparisons among each LPS at 10- and 100-fold excess concentrations, except P > 0.05 for the comparison between msbB LOS and N. meningitidis LOS at a 100-fold excess. Error bars are the standard errors of the means; n = 3.

Article Snippet: Rabbit antiserum to recombinant human LBP (Synpep, Dublin, CA) was purified for immunoglobulins with a HiTrap rProtein A affinity column as per the manufacturer's instructions (Amersham Biosciences).

Techniques: Binding Assay, Concentration Assay, Incubation, Control, Comparison