Detectability of pituitary PRL and GH by immunoassay is increased by thiols and suppressed by divalent cations.
The amount of pituitary PRL detected by RIA in extracts of tissue and of secretory granules was augmented by thiols or EDTA in the extractant, and diminished by divalent cations or cysteamine. GH was affected by thiols and cations similarly, but to a lesser extent, and was not influenced by EDTA or cysteamine. For full immunologic detection of tissue PRL and GH, thiol-dependent mechanisms appear to be required to unmask immunoreactive sites from the poorly immunoreactive oligomeric granule storage forms. Interpretation of studies which rely on measurement of tissue PRL and GH content by RIA, immunoprecipitation, or electrophoretic detection should be reassessed in light of these observations.
D000818:Animals; D002413:Cations, Divalent; D002417:Cattle; D003594:Cytoplasmic Granules / Q000032:analysis / Q000187:drug effects*; D004492:Edetic Acid / Q000494:pharmacology; D013006:Growth Hormone / Q000032:analysis*; D007106:Immune Sera; D007124:Immunoenzyme Techniques; D007700:Kinetics; D010902:Pituitary Gland / Q000032:analysis / Q000187:drug effects*; D011388:Prolactin / Q000032:analysis*; D013438:Sulfhydryl Compounds / Q000494:pharmacology*
D002413:Cations, Divalent; D007106:Immune Sera; D013438:Sulfhydryl Compounds; D011388:Prolactin; D013006:Growth Hormone; D004492:Edetic Acid