Product Protocol: Egf antibody used to evaluate the effects of sialoadenectomy and exogenous egf on taste bud morphology and maintenance (OASA08640)

Specificity: EGF
Product Page: Egf antibody (OASA08640)
Antibody Pair Available: OASA08640 Capture OASA08641 Detection
Experiment Type: Histological methods

After 14 days of EGF replacement, rats were killed by decapitation and their tongues removed. The tongues were cut transversely into anterior (fungiform) and posterior (circumvallate) portions; these were then halved longitudinally. One half of the tongue was fixed in acid alcohol at 4°C and embedded in paraffin for immunofluorescent staining. Half tongues from two sialoadenectomized animals, two shams and two controls were sectioned in a frontal plane at 7 μm and mounted on four sets of slides. Alternative sections were blocked with normal goat serum for 20 min at room temperature and then incubated in primary antibody for 3 h at 37°C. Primary antibodies used were: rabbit anti-rat EGF IgG; rabbit anti-rat TGFα IgG; or sheep anti-rat EGFR IgG (Simms et al., 1991), a kind gift of N. Chegini, Department of Obstetrics and Gynecology, University of Florida. After rinsing in PBS, sections were incubated with appropriate secondary antibodies (goat anti-rat IgG or goat anti-sheep IgG, Biosource, International; diluted 1:100 in 10% normal goat serum in PBS) conjugated to fluorescein for 1 h at 37°C. Negative controls were preformed by substituting the primary antibodies with normal serum. Sections were mounted in glycerol containing an anti-bleach reagent and examined under epifluorescence with a Nikon FXA Photomicroscope.

1. Using standard immunofluorescent staining methods, the distributions of EGF, EGFR and TGFα were examined in fungiform and circumvallate taste buds from SX, SH and control animals. No discernible differences in the distribution or the intensity of staining for EGF, EGFR or TGFα were observed between the dorsal tongue epithelium and papillae of SX, SH or control animals. 2. EGF was diffusely distributed throughout all layers of the stratified epithelium of the dorsal tongue in SX, SH and control animals, although immunostaining for EGF was slightly more intense in the suprabasal layers in the lingual epithelium (data not shown) and was enhanced in the superficial layers of the epithelium lining the circumvallate trench.

1: Morris-Wiman J, Sego R, Brinkley L, Dolce C. The effects of sialoadenectomy and exogenous EGF on taste bud morphology and maintenance. Chem Senses. 2000 Feb;25(1):9-19. PubMed PMID: 10667989.

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