Bbk 120

Sections were deparaffinised, then subjected to antigenic detection of the epitopes with citrate buffer, and an endogenous peroxidase blockade was made with hydrogen peroxidase (3%), a kit (ref: No. BBK 120, Scy Tek, USA) was used to block the endogenous biotin аnd a reagent to block non-specific binding (Superblock, Scy Tek), followed by incubation for 24 hours at 4°C with anti-goat АСЕ -1:300 (sc-12187, Santa Cruz Biotechnology Inc. USA) and monoclonal anti-α smooth muscle actin (A-2547, Sigma) 1:5000 -NRS/TBS/BSA, next incubated with secondary antibody: biotinylated anti-goat (No. AGL015 Scy Tek., USA) for 10 min. The reaction was visualized with 3,3΄-diaminobenzidine tetrachloride (DAB, ScyTek Lab. Inc., USA); counterstaining was performed with Mayer's hematoxylin. As negative controls, sections in which the primary antibodies were replaced by a buffer solution (PBS) were used. Microphotographs were performed with Nikon Microphot SA microscope (Japan), combined with Camedia-5050Z digital camera (Olympus, Japan) at ×100 and ×400 magnification.

Testes were fixed in Bouin's solution for 24 hours. Sections (5 µm thick) of the testes were deparaffinised, then subjected to antigenic detection of the epitopes with citrate buffer, making an endogenous peroxidase blockade with hydrogen peroxidase 3%; a kit (ref: No. BBK 120, Scy Tek, USA) was used to block the endogenous biotin and reagent was used to block the non-specific binding (Superblock, Scy Tek) followed by incubation for 24 hours (at 4°C) with monoclonal mouse antiCRP, SAA, IL4 (1:150, Dako, Denmark), anti-goat АСЕ (1:300, sc 12187, San.Cruz Biot. Inc.,USA), next incubated with secondary antibody: biotinylated anti-rabbit for 10 min at room temperature.
The reaction was visualized with 3,3 diaminobenzidine tetrachloride (DAB, ScyTek Lab. Inc., USA), and counterstaining with Mayer's hematoxylin. The preparations were observed with a light microscope at 400× magnification.