Thomas O. Bruhn

[38] Assay of corticotropin releasing factor

Publisher Summary The corticotropin releasing factor (CRF) can be assayed by a variety of biological and immunological techniques. The chapter describes two methods: an in vitro assay employing primary anterior pituitary cell cultures and a radioimmunoassay (RIA) procedure. The chapter presents the acid extraction method for tissue fragments and a Bond Elute method for aqueous samples for extraction of CRF. The cell culture assay is the exclusive assay used for the isolation and subsequent characterization of ovine CRF and has, thereby, been useful for the detection and quantification of CRF in crude or partially purified extracts. The method has been applied for a wide range of studies investigating the mode of action of CRF and its interactions with other physiological modulators such as glucocorticoids, catecholamines, and neurohypophysial peptides. This method is useful for the assay of both agonist and antagonist analogs of CRF and of highly purified CRF-like peptides of various mammalian and nonmammalian species.