Figure 8.

Specific interaction of HP-NAP with recombinant human TLR2 protein. (A) Binding of TLR2 to HP-NAP as determined by an ELISA-based solid-phase binding assay. The indicated amounts of recombinant human TLR2-10xHis and TLR4-10xHis proteins were incubated with 0.05 μg of HP-NAP coated on an ELISA plate. The binding of TLR2 and TLR4 to HP-NAP was detected by anti-His tag antibody or its isotype control antibody using the ELISA-based solid-phase binding assay as described in Materials and Methods. The results are presented as absorbance at OD_{450 nm}. Data are expressed as mean ± SD of two independent experiments. (B) Specific binding of TLR2 to HP-NAP as determined by competitive binding with Pam₃CSK₄. The indicated amounts of Pam₃CSK₄ (Pam3) and BSA, as a negative control, were incubated with 0.625 μg of the recombinant human TLR2-10xHis protein at 4 °C overnight and then incubated with 0.05 μg of HP-NAP coated on an ELISA plate. The binding of TLR2 to HP-NAP was detected by the ELISA-based solid-phase binding assay as described in A. The results were presented as absorbance at OD_{450 nm}. Data are expressed as mean ± SD of two independent experiments.