Skip to main content
Figure 3 | Reproductive Biology and Endocrinology

Figure 3

From: Fundulus heteroclitus gonadotropins.5: Small scale chromatographic fractionation of pituitary extracts into components with different steroidogenic activities using homologous bioassays

Figure 3

Hydrophobic-interaction chromatography (HIC) of FPE. Approximately 56 pituitaries were extracted twice with 0.5 ml 1.2 M (NH4)2SO4 in 0.1 M sodium phosphate buffer (pH 6.8) and the extracts were combined, centrifugally filtered, and applied to a Phenyl-5-PW column via the 10-ml Superloop. The effluent was monitored for UV-absorbance (A) and a linear gradient of decreasing (NH4)2SO4 concentration was started after the unadsorbed protein eluted from the column. Fractions of 1.0 ml were collected and tested for (B) maturational and (C) steroidogenic activity. Two peaks of E2-generating activity are indicated (a, b). For this experiment only, positive controls (FPE added at a concentration of 0.5 pit. equiv./well) were 50% GVBD with DHP-, T-, and E2-secretion of 1.3, 0.8, and 7.4 ng/well, respectively.

Back to article page

Reproductive Biology and Endocrinology

ISSN: 1477-7827

Contact us