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Additional info for Ecdysone. From Metabolism to Regulation of Gene Expression
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The supernatant was decanted and the radioactivity of the pellet was determined as described above. Luzern, Switzerland) in 2 ml methanol-acetone (1:1, v/v). Homogenates were left overnight at 4 C and then centri fuged (20 min, 15,000 #). -hexane and water (1:1, v/v). The aq. ). , 1982). The methanol was evaporated under a stream of nitrogen, and the residue was redissolved in 500 ^1 phosphate-buffered saline. A 100 //1 aliquot of each extract was subjected to RIA. Accelerated precipitation One hundred microlitres of 2% normal rabbit serum in buffer was added to the assay vials as a carrier globulin, followed by 100//1 of the appropriate second antibody dilution (goat anti-rabbit gamma globulin, 1:10, v/v, as indicated by the manufacturer).
Great Salt Lake strain) at different stages of their vitellogenic cycle. (—) ng ecdysone equivalents g" 1 fresh body wt. (--) ng ecdysone equivalents ml ' haemolymph. Arrows indicate molts; data between brackets correspond to sets of 20 individuals. An approximate time scale is given. The successive stages of each cycle are: OV— = no vitellogenesis, ovary fully transparant; OV + = a few oocytes have started yolk accumulation; OV + = oocytes accumulating yolk are found over the whole length of the ovary; OV + -I- = oocytes opaque due to the presence of lipovitellin and ordered in a thick white double strand; LS = ripe eggs in lateral sacs.
Hoffmann J. , Hetru C. and Luu B. (1984) Ecdysteroids in ovaries and embryos of Locusta migratoria. In Biosynthesis, Metabolism and Mode of Action of Invertebrate Hormones (Edited by Hoffmann J. A. ), pp. 168-180. Springer-Verlag, Berlin. Landureau J. C. and Grellet P. (1972) Nouvelles techniques de culture in vitro de cellules d'insectes et leurs applica tions. r. Acad. Sci. Paris D274, 1372-1376. Lowry O. , Rosebrough N. , Farr A. L. and Randall R. J. (1951) Protein measurement with the Folin phenol reagent.