Vectastain anti rabbit ig abc ap kit

Immunohistochemistry (IHC) for ISAV was performed as previously described [24 (link)]. Briefly, sections of formalin-fixed paraffin-embedded tissue were prepared on poly-lysine-coated slides, dewaxed, and subjected to microwave oven treatment. Rabbit antibody to recombinant nucleoprotein ISAV (ISAV-NP) [25 (link)] was used for virus detection. The Vectastain ABC-AP kit (Vectastain anti-rabbit Ig ABC-AP kit; AK 5001; Vector Laboratories, Inc.) was used for detection of bound antibody employing Fast Red (1 mg mL) and naphthol AS-MX phosphate (0.2 mg mL) with 1 mM levamisole in 0.1 M Tris-buffered saline (TBS) (pH 8.2) as the substrate.

Immunohistochemistry (IHC) for ISAV was performed as previously described [15 (link)] on formalin fixed paraffin embedded gills from farm I and II, and on frozen gills from farm IV. Briefly, polyclonal rabbit antibody to recombinant ISAV nucleoprotein (NP) [16 (link)] was used for the virus detection. Vectastain ABC-AC kit (Vectastain anti rabbit Ig ABC-AP kit, AK 5001, Vector Laboratories, Inc.) was used for detection of bound polyclonal antibody using Fast Red (1 mg ml^-1) and Naphtol AS-MX phosphate (0.2 mg ml^-1) with 1 mM Levamisole in 0.1 M TBS (pH 8.2) as a substrate. In addition, a monoclonal antibody to ISAV haemagglutinin esterase (HE) [17 (link)] was used for the virus detection on the cryosections. An HRP conjugated anti-mouse Ig amplified detection system (EnVision+, Dako) was used for detection of bound monoclonal antibodies using 3,3’-diaminobenzidine (DAB) as substrate. All sections were counterstained with haematoxylin.