Demonstration of "big" CRF (corticotrophin-releasing factor) in bovine hypophyseal stalk.

0.1 N HCl extracts of bovine hypophyseal stalk were fractionated with Sephadex G columns using 0.2 N acetic acid as an eluant. CRF activity of each fraction was assayed with an in vitro system employing cultured rat adenohypophyseal cells. There were 2 discrete CRF peaks with fractionation on G-100, one (Peak A) corresponding to the void volume (MW>150,000). The other (Peak B) was more retarded and eluted slightly in front of immunoreactive ACTH. CRF activity in Peak A was labile during prolonged freezing at low pH. The CRF dose-response slopes for peaks A and B were parallel and were much steeper than that for bovine cerebral cortex extract. Peak A CRF may be a precursor or carrier-bound form of Peak B CRF.