Measurement of activin in biological fluids by radioimmunoassay, utilizing dissociating agents to remove the interference of follistatin.
Activin, a dimer of the beta-subunits of inhibin, is a member of the transforming growth factor beta (TGF-beta) superfamily of growth factors and has a widespread range of actions in a variety of tissues. The investigation of the physiology of activin action has been facilitated in recent years by the availability of immunoassays in addition to bioassays. Follistatin has been shown to bind to activin with a high affinity and therefore interferes in both radioimmunoassays and enzyme-linked immunosorbent assays (ELISAs). In this study we examined the effect of various surfactants and 1.4-dioxane on the measurement of activin in the presence of follistatin by radioimmunoassay. The addition of a combination of sodium deoxycholate. Tween 20 and sodium dodecyl sulphate removed the interference of follistatin in the radioimmunoassay. The measured content of activin in male rat serum, human male serum, human female serum and bovine follicular fluid rose from 3.29 to 4.15, < 0.48 to 2.87, 2.42 to 4.17 and 30.9 to 85.6 ng/ml, respectively, when assayed in the presence of the dissociating reagents. It was unclear whether the altered potencies were due to a dissociation of the follistatin/activin complex rather than the exposure of the epitope on activin recognized by the antiserum. Serum concentrations of activin were lower than those found in testicular cytosols, and after castration no change in serum activin levels was observed, suggesting that the testis does not contribute significantly to circulating activin levels. The use of the dissociating reagents in the radioimmunoassay will enable studies to be carried out that more accurately measure the activin content of various biological fluids, and thus lead to a greater understanding of the physiology of this growth factor.
McFarlane
J R
JR
Institute of Reproduction & Development Monash University, Clayton, Victoria, Australia.
Foulds
L M
LM
Pisciotta
A
A
Robertson
D M
DM
de Kretser
D M
DM
D028341:Activins; D000818:Animals; D001826:Body Fluids / Q000737:chemistry*; D002417:Cattle; D003840:Deoxycholic Acid; D004146:Dioxanes; D005260:Female; D015571:Follicular Fluid / Q000737:chemistry; D038681:Follistatin; D006023:Glycoproteins / Q000302:isolation & purification*; D006801:Humans; D007202:Indicators and Reagents; D007265:Inhibins / Q000032:analysis* / Q000097:blood; D008297:Male; D009919:Orchiectomy; D011136:Polysorbates; D011863:Radioimmunoassay / Q000379:methods*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012016:Reference Values; D012967:Sodium Dodecyl Sulfate; D013501:Surface-Active Agents
D004146:Dioxanes; D038681:Follistatin; D006023:Glycoproteins; D007202:Indicators and Reagents; D011136:Polysorbates; D013501:Surface-Active Agents; D003840:Deoxycholic Acid; D028341:Activins; D012967:Sodium Dodecyl Sulfate; D007265:Inhibins; C025223:1,4-dioxane