Two polyclonal antisera to rat luteal LH/CG receptor with different ligand binding inhibition and immunohistochemical receptor detection capabilities.

https://arctichealth.org/en/permalink/ahliterature57762
Source
Acta Endocrinol (Copenh). 1991 Sep;125(3):305-12
Publication Type
Article
Date
Sep-1991
Author
J T Lakkakorpi
K P Keinänen
H J Rajaniemi
Author Affiliation
Biocenter, University of Oulu, Finland.
Source
Acta Endocrinol (Copenh). 1991 Sep;125(3):305-12
Date
Sep-1991
Language
English
Publication Type
Article
Keywords
Animals
Blotting, Western
Chromatography, Affinity
Electrophoresis, Polyacrylamide Gel
Female
Immune Sera - immunology
Immunohistochemistry
Rats
Rats, Inbred Strains
Receptors, Gonadotropin - immunology - metabolism
Receptors, LH - immunology - metabolism
Research Support, Non-U.S. Gov't
Abstract
Polyclonal antisera to a SDS-denatured and partially renatured rat luteal 90 K LH/CG receptor were raised in rabbits, characterized, and their applicability for immunohistochemical location of the receptor examined. The LH/CG receptor was purified by hCG-affinity chromatography and subjected either to a preparative SDS-PAGE or Western blotting. Gel slices containing the SDS-denatured or nitrocellulose strips containing the renatured 90 K LH/CG receptor were used for immunization. The antisera, termed ARS-2 and ARS-3, respectively, possessed similar antibody titres. Both antisera were able to recognize the native, SDS-denatured, and SDS-denatured and reduced forms of the LH/CG receptor on dot blots, but only ARS-3 contained antibodies to the hormone binding site or a region near to it, as it was able to inhibit the hCG binding to the membrane-bound LH/CG receptor in a dilution-dependent manner. Both antisera recognized the receptor-hCG complex, but ARS-2 stained the complex with about 50% less intensity than the free receptor. ARS-3 located the LH/CG receptor distinctly on the luteal cell surfaces in immunohistochemical staining with peroxidase antiperoxidase complex method, but ARS-2, although it possessed similar antibody titre, revealed negligible staining. Thus, the antisera readily recognize the native receptor, but differ in their capability for inhibiting hormone binding. Only ARS-3, produced against the renatured receptor, contains sufficient amounts of antibodies capable of recognizing free and occupied receptors in immunohistochemistry.
PubMed ID
1950343 View in PubMed
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