Determination of locust vitellogenin by radioimmunoassay with [3H]Propionyl-vitellogenin.
A simple procedure for radiolabeling of locust vitellogenin is described. This procedure involves coupling of [3H]propionyl succinimidate to purified vitellogenin with high yield and specific activity. Using this radiolabeled analog, a specific and sensitive radioimmunoassay was developed for determining locust vitellogenin content, with a lower detection limit of 1 ng. [3H]Propionyl-vitellogenin binds completely to rabbit anti-vitellogenin (locust) and can be completely competed out by locust vitellogenin. The structural similarity of locust vitellogenin with that of locust egg vitellin, male locust lipophorin (a diglyceride-carrying lipoprotein), Xenopus laevis vitellogenin, and chicken egg yolk lipovitellin was examined with this RIA procedure. Comparable binding competition was obtained with locust vitellin only. Male locust lipophorin, Xenopus vitellogenin, and chicken lipovitellin did not inhibit vitellogenin binding at concentrations 1000-fold greater than that of locust vitellogenin. The use of this RIA in determination of vitellogenin synthesis in vivo and in vitro, using isolated fat body preparations, is described.
D000818:Animals; D005216:Fat Body / Q000032:analysis; D006110:Grasshoppers / Q000378:metabolism*; D006458:Hemolymph / Q000032:analysis; D008074:Lipoproteins / Q000032:analysis*; D011863:Radioimmunoassay; D014818:Vitellogenesis; D014819:Vitellogenins / Q000031:analogs & derivatives / Q000032:analysis*
D008074:Lipoproteins; D014819:Vitellogenins; C039994:propionylvitellogenin