Comparison of immunoreaction between two commercial antibodies against 14-3-3σ protein in canine tissues

Figures & data

Table 1. Reactivity patterns of polyclonal (N14) and monoclonal (5D7) antibody to 14-3-3σ in normal canine tissues.

Tissue	N14^a	5D7^a
Skin
Epidermal/hair follicles	+++	+++
Sebaceous gland	++	++
Apocrine glands
Acini	++	++
Ducts	++	++
Digestive system
Squamous epithelium (oral cavity)	+++	+++
Salivary glands
Acini	−	−
Myoepithelium	−	−
Ducts	−	−
Oesophagus
Surface epithelium	+	+
Glands	+	+
Stomach
Gastric pits	+	−
Corpus glands	−	−
Pyloric glands	−	−
Neuroendocrine cells	+	−
Intestine
Surface epithelium	−	−
Mucosal crypts	−	−
Liver
Hepatocytes	−	−
Bile ducts	−	−
Gall bladder	+++	+++
Pancreas
Acini	−	−
Ducts	−	−
Islet	+++	−
Respiratory tract
Trachea
Cylindrical ciliated epithelium	−	−
Basal cells	+	+
Bronchioles
Cylindrical ciliated epithelium	−	−
Basal cells	+	+
Alveolar epithelium	−	−
Glands	−	−
Urinary system
Kidney
Bowman’s capsule	−	−
Collecting ducts	−	−
Urinary bladder
Superficial cells	+++	+++
Basal cells	+++	+++
Endocrine glands
Pituitary glands	−	−
Thyroid/parathyroid	−	−
Adrenal gland
Cortex	−	−
Medulla	++	−
Male genital
Seminiferous tubules	−	−
Excretory ducts	−	−
Epididymis	−	−
Prostate	++	++
Female genital
Ovary
Surface epithelium	−	−
Rete ovarii	−	−
Endometrium
Surface epithelium	−	−
Glands	+	−
Squamous epithelium (vagina)	+++	+++
Mammary glands
Secretory cells	+/−	+/−
Myoepithelium	+++	+++
Ducts	+/−	+/−
Nervous system
Cerebrum	−	−
Cerebellum	−	−
Ependymal cells	−	−
Ganglia	+	−

^a −, No stained cells; +, faint; ++, medium or heterogeneous; +++, strong reaction; +/−, solitary positive cells.

Figure 1. 14-3-3σ immunohistochemistry: in canine adrenal gland – (a) note strong cytoplasmic immunoreaction in cells of the medulla when the polyclonal antibody is employed and (b) the incubation with clone 5D7 avoids this reaction; in canine pancreas – (c) N14 showed immunoreaction and (d) 5D7 do not detect 14-3-3σ in islet of Langerhans; in canine gastric mucosa – (e) supranuclear granular staining in both parietal and zymogenic cells with N14 and (f) canine stomach do not produce any immunoreaction with monoclonal antibody 5D7.

Figure 2. Western blot analysis: (1) mammary gland, (2) urinary bladder, (3) salivary gland, (4) oesophagus, (5) skin, (6) kidney, (7) cerebrum, (8) cerebellum and (9) Lymph node.