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2 protocols using purified anti human tlr4 antibody

1

Investigating TLR4 and Mac-1 Activation

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KLA (Kdo-Lipid A) was purchased from Adipogen (San Diego, CA, USA). Purified anti-human TLR4 antibody, anti-human Mac-1 antibody and anti-mouse IgG isotype control antibody were purchased from eBioscience (San Diego, CA). R-phycoerythrin (PE)-conjugated mouse anti-human Mac-1 (clone ICRF44; BD) antibody and PE-conjugated mouse IgG isotype control (clone MOPC-21) antibody were obtained from BD (San Diego, CA). Rabbit anti-mouse Mac-2 antibody was purchased from Abcam (Cambridge UK). MK-886 and various signal pathway inhibitors were purchased from EMD Serono, Inc. (Rockland, MA, USA) and Sigma-Aldrich Co. (Saint Louis, MO).
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2

Escherichia coli LPS Signaling Pathway

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LPS from Escherichia coli was purchased from Sigma-Aldrich (Saint Louis, MO). pGL3 basic vector, pRL CMV vector, and dual luciferase reporter assay kits were purchased from Promega (Madison, WI). DNeasy Tissue Kits and QIAprep Spin Kits were supplied by Qiagen (GmhH, Germany). The various signal pathway inhibitors used were acquired from Calbiochem (Ra Jolla, CA) and Sigma (St. Louis, MO). 5-LO antibody were purchased from Santa Cruz Biotechnology (Beverly, MA). Akt, phosphospecific antibody against Akt and IKK were from Cell Signaling Technology (Beverly, MA). Purified anti-human TLR4 antibody was from eBioscience (San Diego, CA). Horseradish peroxidase (HRP)-conjugated IgG (Santa Cruz Biotechnology, Santa Cruz, MA) was used as the secondary antibody.
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