Localization of EGF receptors in frozen tissue sections by antibody and biotinylated EGF-based techniques.
We developed a sensitive EGF receptor detection method for frozen tissue sections using biotinylated EGF as the primary reagent. The method was directly compared with an immunohistochemical technique based on an anti-EGF receptor monoclonal antibody (MAb EGFR1) in normal human and rat tissues and in human tumors. The method was more sensitive than a previously published biotinylated EGF-based technique. In normal human tissues and in 37 of the 50 tumors, the binding pattern mirrored that of positive staining with EGFR1. Five further tumors showed weak immunoreactivity, but in these no binding of biotinylated EGF was detected. The remaining eight tumors were negative by both techniques. The discordant cases may reflect a lower level of sensitivity of the ligand-binding technique or, alternatively, abnormal receptors may have been expressed in these tissues. EGF receptors could be detected in rat liver with biotinylated EGF but not with the antibody, indicating the usefulness of the ligand-based technique in cross-species studies.
Reeves
J R
JR
Department of Surgery, University of Glasgow, Glasgow Royal Infirmary, United Kingdom.
Cooke
T G
TG
Fenton-Lee
D
D
McNicol
A M
AM
Ozanne
B W
BW
Richards
R C
RC
Walsh
A
A
D000818:Animals; D000911:Antibodies, Monoclonal; D001710:Biotin; D004815:Epidermal Growth Factor / Q000032:analysis*; D066246:ErbB Receptors / Q000032:analysis* / Q000276:immunology; D005260:Female; D005629:Frozen Sections; D006801:Humans; D007124:Immunoenzyme Techniques; D008024:Ligands; D008099:Liver / Q000737:chemistry; D008297:Male; D051379:Mice; D008867:Microtomy; D009369:Neoplasms / Q000737:chemistry; D051381:Rats; D012680:Sensitivity and Specificity; D012867:Skin / Q000737:chemistry
D000911:Antibodies, Monoclonal; D008024:Ligands; D004815:Epidermal Growth Factor; D001710:Biotin; D066246:ErbB Receptors