Detection and purification of inhibin using antisera generated against synthetic peptide fragments

Substratum-growth factor collaborations are required for the mitogenic activities of activin and FGF on embryonal carcinoma cells.

“…Although bFGF was isolated as a mitogen (Baird and Bohlen, 1990), activin A was initially purified on the ba- sis of its ability to stimulate FSH secretion (Vaughan et al, 1988) . The only report that activin A can act as a mitogen is with BALB/c 3T3 cells (Kojima and Ogata, 1989) .…”

“…Falcon plastic tissue culture dishes (Falcon Labware, Becton, Dickson and Co., Oxnard, CA) were used exclusively. Recombinant bovine bFGF was a generous gift from Dr. Andrew Baird (Whittier Institute, La Jolla, CA) and porcine activin A was obtained from Joan Vaughan and Wylie Vale (Salk Institute ; see Vaughan et al ., 1988 for purification procedures) .…”

Immunolocalization of Inhibin/Activin Subunit Proteins During the Breeding Season in Testes and Scented Glands of Muskrats (Ondatra zibethicus)

“…The sec-tions were then incubated with primary antibody (1:1,000 or 1:2,000) raised against porcine inhibin α chain (1-30)-NH 2 conjugated to rabbit serum albumin, porcine inhibin/activin  A (81-113)-NH 2 (#305-24D) [30] and cyclic acetyl human inhibin/activin  B (81-113)-NH 2 (#305-25D) [30] for 12 hr at room temperature. The sections were then incubated with a second antibody, goat anti-rabbit lgG conjugated with biotin and peroxidase with avidin, using a rabbit ExtrAvidin TM staining kit (Sigma, St. Louis, MO, U.S.A.), followed by visualizing with 30 mg 3,3-diaminobenzidine (Wako, Tokyo, Japan) solution in 150 ml of 0.05 mol Tris-HCl l -1 buffer, pH 7.6, plus 30 μl H 2 O 2 .…”

“…The membranes were blocked in 3% BSA for 1 hr at room temperature. Primary incubation of the membranes was carried out using a 1:1,000 dilution of primary antibody raised against porcine inhibin α chain (1-30)-NH 2 conjugated to rabbit serum albumin, porcine inhibin/activin  A (81-113)-NH 2 (#305-24D) [30] and cyclic acetyl human inhibin/activin  B (81-113)-NH 2 (#305-25D) [30] for 60 min. Secondary incubation of the membranes was then carried out using a 1:1,000 dilution of goat anti-rabbit IgG tagged with horseradish peroxidase for 60 min.…”

Immunolocalization of Inhibin/Activin Subunits and Steroidogenic Enzymes in Equine Neonatal Ovaries

“…The sections were then incubated with primary antibody (1:8,000) raised against porcine inhibin α chain (1-30)-NH 2 conjugated to rabbit serum albumin, porcine inhibin/activin β A (81-113)-NH 2 (#305-24D) [29], human inhibin/activin β B (81-113)-NH 2 (#305-25D) [29], bovine adrenal cholesterol side-chain cleavage cytochrome P450 (P450scc) [3], human placental 3β-hydroxysteroid dehydrogenase (3βHSD) [7], porcine testicular 17α-hydroxylase cytochrome P450 (P450c17) [12] and human placental aromatase cytochrome P450 (P450arom) [16] for 12 hr at room temperature. The inhibin α subunit peptide was kindly provided by Dr. N. …”