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1992) observed increased IGFBP-2 by immunoblotting media from rat hepatocytes treated with retinoic acid, whereas no change in -32-kDa IGFBF’s was observed by ligand blotting. Specific identification of IGFBPs requires immunologic identification or protein sequencing. Bands staining positively on immunoblots can be directly compared with bands identified by ligand blotting. , 1989a; Yang and Rechler, 1993) provides sensitive assays. , 1992) and IGFBP-6 (Baxter and Saunders, 1992) have recently been described.
Similar results were obtained by Ballard et al. (1991). The alternative possibility that radiolabeled IGF-I1binds to a preexisting ternary complex containing endogenous IGF-I, which would imply that IGFBP-3 has separate binding sites for IGF-I and IGF-11, cannot be formally excluded. However, competitive binding data suggest that IGF-I and IGF-I1 tracers bind to a single site on purified rat IGFBP-3 since binding of both tracers is completely inhibited by unlabeled IGF-I or IGF-I1 (Baxter and Martin, 1987).
Perdue binds normally to recombinant IGFBP-3 (L. A. Bach, unpublished results)]. 7. Site of Synthesis Scott and Baxter (1991) demonstrated that cultured rat hepatocytes synthesized ALS activity. ALS activity was stimulated twofold when hepatocytes were incubated with bovine GH. , 1992). 8. Properties of Rat ALS Several observations suggest that unlike human ALS, rat ALS may bind free IGFBP-3 as well as IGF-I (IGF-II):IGFBP-3binary complexes. Yang et al. (1989)demonstrated that after fractionation of adult rat serum, IGF-I1tracer bound to presumably unoccupied binding sites in the 150-kDa region; these sites had preferential affinity for IGF-11.