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Pam3csk4 tlr1 2

Manufactured by InvivoGen
Sourced in United States

Pam3CSK4 (TLR1/2) is a synthetic triacylated lipopeptide that serves as a potent agonist for the Toll-like receptor 1/2 (TLR1/2) complex. It can be used as a research tool to study the activation of the TLR1/2 signaling pathway.

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3 protocols using pam3csk4 tlr1 2

1

Toll-like Receptor Signaling in Inflammation

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Pam3CSK4 (TLR1/2) and LPS (TLR4) were obtained from InvivoGen (Carlsbad, CA, USA). Mouse recombinant IFN-β was purchased from PBL InterferonSource (now PBL Assay Science, Piscataway, NJ, USA) and mouse IFN-γ was from R&D Systems (Minneapolis, MN, USA). Antibodies used in this study were obtained as follows. Anti-H-PGDS and Anti-COX-2 antibodies were produced by Cayman Chemical (Ann Arbor, MI, USA). Anti-STAT1 and anti-pY701-STAT1 antibodies were from Cell Signaling Technology (Danvers, MA, USA). Anti-actin antibody was from BD Biosciences (Franklin Lakes, NJ, USA). PGs were purchased from Cayman Chemical (Ann Arbor, MI, USA). All other chemicals were from Sigma (St. Louis, MO, USA), unless stated otherwise.
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2

Immune Modulation by TLR Agonists and Anti-inflammatory Drugs

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TLR agonists LPS (TLR4), R848 (TLR7/8) and Pam3CSK4 (TLR1/2) (all from Invivogen, San Diego, California, USA) were used at a final concentration of 2 µg/mL, as described previously.77 (link) In addition to these TLR agonists, the following anti-inflammatory drugs were used in this study: IBF and BMS (both from Sigma-Aldrich). IBF was resuspended in sterile PBS, while BMS was initially resuspended in 100% ethanol before diluting 1:5 with sterile PBS, and both drugs were used at a final concentration of 1 µg/mL, which was the drug concentration previously optimized in anti-inflammatory drug titration experiments (data not shown).
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3

Optimizing TLR Agonists for HIV Infection

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The TLR agonists LPS (TLR4), R848 (TLR7/8), and Pam3CSK4 (TLR1/2) (all from Invivogen, San Diego, CA, USA) concentrations were previously optimized in TLR titration experiments. As no significant differences were observed in HIV infections (Supplementary Figure 1) or cytokine profiles (Supplementary Figures 24) between the TLR concentrations, a final concentration of 2 μg/ml was used. Phytohaemagglutinin (PHA) (Sigma-Aldrich, St. Louis, MO, USA), used as the positive control at a final concentration of 10μg/ml. Unstimulated PBMCs were used as the negative control. The CCR5-tropic HIV-1 NL4-3 AD8 (35 (link)) (a gift from Dr. Alex Sigal), was used at a working dilution of 1:20, which corresponded to an MOI of 0.9, which had been previously optimized (data not shown). PHA and unstimulated uninfected conditions were used as controls for HIV.
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