Reagents:
… Ultrapure LPS from Salmonella minnesota R595 (Re) and Ultrapure LPS from Escherichia coli 055:B5 were obtained from List Laboratories. …
Human Macrophage Assay with LPS and Nickel:
… Five days later, the differentiated macrophages were incubated with a dose range of Hu 15C1 together with ultrapure LPS from S. minnesota (1, 10, or 100 ng/ml; List Biological Laboratories)…
U937 Cell Assays:
… The cells were subsequently incubated with 1 ng/ml ultrapure LPS from S. minnesota for 20 h, and IL-6 in the supernatants was quantified using the Quantikine ELISA kit (R&D Systems).
Confocal Microscopy and FRET Analysis:
THP-1 cells either untreated or treated with ultrapure LPS from E. coli (100 ng/ml) in the presence or absence of different concentrations (0.1, 1, or 10 g/ml) of Hu 15C1…
THP-1 Blue Prolonged Blockade Assay:
… The cells were resuspended in fresh medium and incubated for 0, 1, or 24 h prior to stimulation with 5 ng/ml ultrapure LPS from S. minnesota for 24 h. NF-B reporter gene activation, …
LPS Lung Instillation:
C57BL/6 mice (female, 8 weeks old) from Charles River Laboratories were injected with the indicated antibodies intravenously. One hour later, mice were anesthetized and instilled intranasally (i.n.) with 10 g of ultrapure LPS from S. minnesota. 24 h post-LPS challenge, mice were sacrificed, and bronchoalveolar lavage (BAL) was performed…