Homologous somatotropin radioreceptor assay utilizing recombinant bovine growth hormone.
A homologous radioreceptor assay using recombinant bovine growth hormone and bovine liver membranes is described. The total specific binding of 125I-labeled recombinant bovine growth hormone to the 100 000 X g pellet was 48% in 24 h at 25 degrees C. Hormone binding was partially reversible, with 40% of the radiolabeled hormone being irreversibly bound. The amount of specific binding varied with assay pH, with the optimum occurring at pH 7.8. Specific binding was temperature-dependent, with greater specific binding occurring at 25 degrees C than at 5 degrees C or 37 degrees C during a 24 h period. Recombinant bovine growth hormone, human growth hormone, ovine growth hormone and recombinant porcine growth hormone competed effectively with 125I-labeled recombinant bovine growth hormone for binding sites, while bovine prolactin and ovine prolactin were needed in amounts 10(6)-fold the concentration of recombinant bovine growth hormone to displace the radiolabeled hormone. Surprisingly, human placental lactogen did not displace the radiolabeled hormone.
D000818:Animals; D002417:Cattle; D013006:Growth Hormone; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D007457:Iodine Radioisotopes; D007700:Kinetics; D008566:Membranes / Q000378:metabolism; D008862:Microsomes, Liver / Q000378:metabolism; D010928:Placental Lactogen / Q000494:pharmacology; D011388:Prolactin / Q000494:pharmacology; D011506:Proteins / Q000378:metabolism; D011869:Radioligand Assay / Q000379:methods; D011956:Receptors, Cell Surface / Q000378:metabolism*; D011986:Receptors, Somatotropin; D013696:Temperature; D013997:Time Factors
D007457:Iodine Radioisotopes; D011506:Proteins; D011956:Receptors, Cell Surface; D011986:Receptors, Somatotropin; D011388:Prolactin; D013006:Growth Hormone; D010928:Placental Lactogen