Immunohistochemistry Protocol for Feline COX-1, COX-2, and CD147

Immunohistochemistry protocols (IHC) were designed for the detection of cyclooxygenase-1 (COX-1), COX-2, and CD147. The COX-1 antibody was a polyclonal rabbit anti-ovine COX-1 antiserum (Cayman Chemical Company, Ann Arbor, MI, USA). The other antibodies were polyclonal rabbit anti-mouse COX-2 IgG (Cayman Chemical) and polyclonal goat anti-human CD147 IgG (Santa Cruz Biotechnology Inc., Dallas, TX, USA). Comparison of epitope sequence and predicted feline amino acid sequences available in NCBI databases revealed that the COX-2 antibody recognized an epitope that was 83% identical to feline COX-2, and the COX-1 antiserum recognized an epitope that was 91% identical to feline COX-1. The epitope sequence for the CD147 antibody was proprietary, but personal communication with the manufacturer (Santa Cruz) indicated 79% homology to feline CD147. Feline intestinal goblet cells were selected as a positive control for COX-1 [29 (link)], feline kidney (macula densa) was selected as a positive control tissue for COX-2 [30 (link)], and feline small intestine was selected as a positive control for CD147 expression [31 (link)]. For negative control purposes, primary antibodies and antiserum were replaced with control rabbit IgG (Millipore Sigma Company, Etobicoke, ON, Canada), control goat IgG, and control rabbit serum (Santa Cruz).

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Nasry W.H., Wang H., Jones K., Dirksen W.P., Rosol T.J., Rodriguez-Lecompte J.C, & Martin C.K. (2018). CD147 and Cyclooxygenase Expression in Feline Oral Squamous Cell Carcinoma. Veterinary Sciences, 5(3), 72.

Publication 2018

control goat IgG

Amino acid sequences Anti igg Antibodies Antibody Antiserum Cayman Cd147 Cox 2 Cyclooxygenase 1 Epitope Feline Goat Goblet cells Human Immunohistochemistry Intestinal Kidney Macula Mouse Ovine Rabbit Serum Small intestine Tissue

Corresponding Organization : University of Prince Edward Island

Other organizations : The Ohio State University, Ohio University

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Variable analysis

independent variables

Type of antibody used for immunohistochemistry (COX-1, COX-2, CD147)

dependent variables

Detection (presence/absence) of COX-1, COX-2, and CD147 proteins in feline tissues

control variables

Epitope sequence homology between antibodies and feline proteins (COX-1, COX-2, CD147)
Positive control tissues (feline intestinal goblet cells for COX-1, feline kidney macula densa for COX-2, feline small intestine for CD147)
Negative control antibodies (control rabbit IgG, control goat IgG, control rabbit serum)

controls

Feline intestinal goblet cells for COX-1, Feline kidney (macula densa) for COX-2, Feline small intestine for CD147
Control rabbit IgG, Control goat IgG, Control rabbit serum

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