Localization of EGF receptors in frozen tissue sections by antibody and biotinylated EGF-based techniques.

Reeves, J R; Cooke, Timothy G.; Fenton-Lee, D; McNicol, A. M.; Ozanne, Brad; Richards, R. C.; Walsh, Alex J.

doi:10.1177/42.3.8308248

Cited by 14 publications

(12 citation statements)

References 2 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…EGFRI was detected using biotinylated rabbit anti-mouse antibody, and non-immune mouse IgG2b was used as a control for non-specific antibody binding. (Reeves et al, 1993).…”

Section: Cellmentioning

confidence: 99%

See 1 more Smart Citation

Epidermal growth factor receptor expression by human squamous cell carcinomas of the head and neck, cell lines and xenografts

Stanton

Richards²,

Reeves³

et al. 1994

Br J Cancer

View full text Add to dashboard Cite

show abstract

“…EGFRI was detected using biotinylated rabbit anti-mouse antibody, and non-immune mouse IgG2b was used as a control for non-specific antibody binding. (Reeves et al, 1993).…”

Section: Cellmentioning

confidence: 99%

“…Human syncytiotrophoblast plasma membranes were prepared by cold isotonic saline extraction (Smith et al, 1974 (Reeves et al, 1993). EGFRI was detected using biotinylated rabbit anti-mouse antibody, and non-immune mouse IgG2b was used as a control for non-specific antibody binding.…”

Section: Cellmentioning

confidence: 99%

Epidermal growth factor receptor expression by human squamous cell carcinomas of the head and neck, cell lines and xenografts

Stanton

Richards²,

Reeves³

et al. 1994

Br J Cancer

View full text Add to dashboard Cite

show abstract

“…With some previously reported protocols, no binding could be detected in tissues with moderate levels of receptors (Reeves et al 1994). In this case, the in situ IGF-1-His binding gave an overall staining pattern well representing the known IGFR-1 distribution.…”

Section: Discussionmentioning

confidence: 71%

“…Some techniques uses concentrations of ligand between 8-and 1000-fold greater than the dissociation constant of the ligand-receptor interaction (Michel and Parsons 1988,1990Zhang et al 1993;Bentham et al 1994;Ahmed et al 1995). For near-maximal saturation of the receptors, there should be no need to use concentrations of the ligand that are more than about fivefold greater than the dissociation constant of the ligand-receptor interaction (Reeves et al 1994). Reported values for the IGFR-1 are in the range of 0.1-1 nM (Siddle and Soos 1999).…”

Section: Discussionmentioning

confidence: 99%

“…Recently, methods using fluorescently labeled or biotinylated ligands were described for localization of receptors in tissue sections Parsons 1989,1990;Reeves et al 1994;Göritz et al 1996;Ray and Ariano 1998). Binding of fluorophore-conjugated ligands is directly visualized using a fluorescent microscope, whereas the biotinylated ligands are detected using a streptavidin-conjugated enzyme and a chromogenic substrate.…”

mentioning

confidence: 99%

See 1 more Smart Citation

Novel Non-isotopic Method for the Localization of Receptors in Tissue Sections

Desnoyers¹,

Simonette²,

Vandlen³

et al. 2001

J Histochem Cytochem.

View full text Add to dashboard Cite

S U M M A R YWe describe a novel fluorescent method for the detection of receptors for chimeric proteins in tissue sections. The technique was developed using a recombinant human insulin-like growth factor (IGF-1) chimera, bearing six additional histidine residues at the carboxy-terminal end (IGF-1-His). We demonstrated that dehydration of the tissue sections was detrimental for binding and that its prevention dramatically increased sensitivity. The specificity of IGF-1-His interaction was shown by gradual abolition of the fluorescent signal in the presence of increasing concentrations of IGF-1. Combining immunofluorescence with in situ ligand binding, we showed that IGF-1-His binding corresponded to the IGF-1 receptor (IGFR-1) distribution in human fetal kidney. Moreover, incubation of the tissue sections with an anti-IGFR-1 blocking antibody abolished IGF-1-His binding, demonstrating that the interaction was mediated by the IGFR-1. The method was also used to localize the IGFR-1 in E18 rat embryo sagittal sections. The IGF-1-His binding pattern was observed in brain, cartilage, lung, skin, heart, diaphragm, and tongue, and paralleled the previously reported IGFR-1 distribution. We believe that this new non-isotopic in situ ligand binding method will facilitate rapid and accurate localization of receptors in tissue sections.

show abstract

Two lectin-like receptors for α1-acid glycoprotein in mouse testis

1996

View full text Add to dashboard Cite

Three glycoforms of alpha 1-acid glycoprotein (AGP) were biotinylated to examine their binding in mouse testis by light microscopy. The transition from one stage to another in the spermatogenic cycle is marked with an appearance of a receptor for the Concanavalin A (Con A) non-reactive glycoform AGP-A in the cytoplasm of spermatocytes, young spermatids and Sertoli cells. This receptor disappears in the late stages of the spermatids. The Con-A intermediately reactive and the Con-A reactive glycoforms, AGP-B and AGP-C, showed weak reaction in the cytoplasm of spermatocytes, spermatids and Sertoli cells and, at the last stages in the spermatogenic cycle, a very strong reaction in the late elongated spermatids and the apical extensions of Sertoli cells. The interactions are lectin-like as confirmed by inhibition with simple sugars. In addition, the bindings were inhibited by steroid hormones. AGP-A was inhibited by testosterone, oestradiol and progesterone, while AGP-B and AGP-C were inhibited by mannose, GlcNAc, cortisone, aldosterone, oestradiol and progesterone. The receptors and the corresponding AGP glycoforms may be adhesion molecules between Sertoli cells and the spermatogenic cells and may have a function as a regulatory factor for spermatozoa development.

show abstract

Localization of EGF receptors in frozen tissue sections by antibody and biotinylated EGF-based techniques.

Cited by 14 publications

References 2 publications

Epidermal growth factor receptor expression by human squamous cell carcinomas of the head and neck, cell lines and xenografts

Epidermal growth factor receptor expression by human squamous cell carcinomas of the head and neck, cell lines and xenografts

Novel Non-isotopic Method for the Localization of Receptors in Tissue Sections

Two lectin-like receptors for α1-acid glycoprotein in mouse testis

Contact Info

Product

Resources

About