Toll-like receptor 2 blocking experiments - (Apr/24/2006 )

I'm trying to block TLR-2 agonist induced cytokine production from human PBMCs with the TLR-2 antagonist TLR2.1 (eBioscience).
Before stimulation with PamCys or LTA I pre-incubate my cells for 30 min at room temperature with 10 microgram/mililiter TLR2.1
Unfortunately it doesn't work, even when I double the concentration of my TLR-2 antagonist.
Does anybody know what I'm doing wrong?

Thanx,

Marcel

we use a TLR2 blocking antibody sometimes (with LTA as one of the ligands) with human epithelial cells...we add antibody diluted in media, put the cells back in the 37 degree CO2 incubator for an hour, then treat with our stimulant, while maintaining the TLR2 antibody at the same concentration during treatment...works pretty well