Ileal tissue sections were prepared and stained as described in the materials and methods. Briefly, tissue sections were treated with Dako target retrieval solution for unmasking the tissue antigens. After blocking, tissue sections were incubated with goat antihuman TLR-2, -4, -5 and -9 antibodies. Normal goat IgG treatment served as an isotype antibody negative control. Folllowing 3X washings, tissue sections were incubated with HRP-conjugated rabbit anti-goat IgG antibody, treated with DAB and counter-stained with Mayer’s hematoxylin. The sections were dehydrated and mounted permanently. The specificity of each TLR antibody was tested by pre-incubating TLR antibodies with the corresponding peptides and this treatment completely abrogated the TLR staining. The tissue sections from various treatment groups showed TLR-2 and TLR-9 specific staining. No TLR-4 and -5 specific tissue staining was observed.
Figure 2: Detection of TLR-2 and TLR-9 proteins on porcine ileal tissues by immunohistochemical (IHC) staining.