Technical note: p40 antibody as a replacement for p63 antibody in bovine mammary immunohistochemistry

• Published in: Journal of dairy science Vol. 101; no. 8; pp. 7614 - 7617
• Main Authors: Parsons, C.L.M., Tucker, H.L.M., Akers, R.M., Daniels, K.M.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.08.2018
• Subjects: Animals; Antibodies, Monoclonal; Cattle; Female; Immunohistochemistry - methods; Immunohistochemistry - veterinary; Lactation; mammary; myoepithelial; p40; p63; Protein Isoforms; Tumor Suppressor Proteins - analysis; Tumor Suppressor Proteins - metabolism; myoepithelial; mammary; p63; p40
• ISSN: 0022-0302; 1525-3198
• DOI: 10.3168/jds.2018-14676

Tumor protein 63 (p63) is a nuclear antigen found in basal epithelial cells. To date, 10 isoforms of p63 have been identified, falling into 2 major groups identified by presence or absence of an N-terminal transactivation domain (TAp63 and ΔNp63, respectively). Literature suggests that ΔNp63 is the predominant form expressed in basal epithelial cells and myoepithelial cells (MYEC). The mouse anti-p63 antibody, clone 4B1E12, has been used as a specific nuclear marker for bovine MYEC. Unfortunately, this antibody is no longer commercially available. A new mouse monoclonal antibody, clone BC28, specific to ΔNp63 (designated p40) has been developed. We hypothesized that the p40 antibody would be an appropriate substitution as a MYEC and epithelial basal cell marker. An array of archived formalin-fixed, paraffin-embedded bovine tissues were subjected to immunohistochemical staining for either p40 or p63, with a subset being dual stained for direct comparison. Positive staining for p40 and p63 was observed in serial sections of mammary, skin, rumen, salivary gland, ureter, and bladder. As predicted, negative staining for p40 and p63 was observed in testis and intestine. Dual staining for p40 and p63 in calf mammary (n = 4), lactating mammary (n = 4), rumen (n = 4), and skin (n = 4) showed nearly 100% agreement. Thus, we established that the mouse monoclonal antibody, clone BC28, is a suitable replacement for anti-p63, clone 4B1E12, as a marker of MYEC and basal epithelial cells in bovine tissues.