Immunohistochemical detection of human estrogen receptor with region D-specific antipeptide antibodies

• Published in: The Journal of steroid biochemistry and molecular biology Vol. 43; no. 4; pp. 311 - 317
• Main Authors: Poutanen, M., Mäentausta, O., Isomaa, V., Jouppila, P., Vihko, R.
• Format: Journal Article
• Language: English
• Published: Oxford Elsevier Ltd 01.10.1992; Elsevier Science
• Subjects: Biological and medical sciences; Breast Neoplasms - metabolism; Cell receptors; Cell structures and functions; Endometrium - metabolism; Female; Fundamental and applied biological sciences. Psychology; Hormone receptors. Growth factor receptors. Cytokine receptors. Prostaglandin receptors; Humans; Immunohistochemistry; Molecular and cellular biology; Peptides - immunology; Precipitin Tests; Receptors, Estrogen - analysis; Receptors, Estrogen - metabolism; Human; Immunohistochemistry; Antibody; Peptide fragment; Estrogen; Sex steroid hormone; Polyclonal immunoglobulin; Hormonal receptor
• ISSN: 0960-0760; 1879-1220
• DOI: 10.1016/0960-0760(92)90166-G

To study the possibility of using antipeptide antibodies for the immunohistochemical determination of human estrogen receptors (ER), three peptides corresponding to the putative major antigenic regions of the human ER (Met 12-Leu 26, or ERP1; Thr 227-Gln 267, or ERP2; Leu 256-Gly 275, or ERP3) were used to produce site-specific rabbit polyclonal antipeptide antisera. High titer antibodies were obtained against all the peptides used, as judged by time-resolved fluoroimmunoassay. The antibodies against region D (ERP3) specifically immunoprecipitated the ER proteins in vitro, as did the antiERP2 antibodies to a much smaller extent. With one of the region D-specific antibodies (antiERP3 Ab2) ER could also be immunohistochemically detected. When benign and malignant human breast and normal endometrial tissues were used, the immunohistochemical staining observed with these antipeptide antibodies correlated well with the staining obtained with an established method. Thus, the results reported here show that this part of region D in ER is a potential antigenic epitope for the production of site-specific antibodies against ER. Antipeptide antibodies produced against this region can be used to immunolocalize the ER in various normal and pathological human tissues.