THP1-Blue™-CD14 cells are a human monocytic cell line engineered to stably express a secreted embryonic alkaline phosphatase (SEAP) reporter gene under the control of a NF-κB-inducible promoter. The cells constitutively express CD14, a key receptor involved in the recognition of lipopolysaccharide (LPS) and other microbial components.
Example 6
THP1-Blue™-CD14 cells (Invivogen), which express human TLR4/MD2 complex, were plated in 96 well plates at 105 cells/well in 30 μl of HEK-Blue detection medium (Invivogen). TLR4 antibodies were diluted in 30 μl of medium to the appropriate concentration and added to the cells for 30 min at 37° C. Then, LPS was diluted at 10 ng/mL in 30 μl of medium, added to the cells, and left to incubate 24 h at 37° C. The absorbance was measured at 650 nm using a microplate reader.
Example 6
THP1-Blue™-CD14 cells (Invivogen), which express human TLR4/MD2 complex, were plated in 96 well plates at 105 cells/well in 30 μl of HEK-Blue detection medium (Invivogen). TLR4 antibodies were diluted in 30 μl of medium to the appropriate concentration and added to the cells for 30 min at 37° C. Then, LPS was diluted at 10 ng/mL in 30 μl of medium, added to the cells, and left to incubate 24h at 37° C. The absorbance was measured at 650 nm using a microplate reader.
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