Growth and Differentiation Factor 9, Recombinant, Mouse (GDF9)

Mouse GDF-9 Growth differentiation factor-9 (GDF-9) is a member of the transforming growth factor-beta (TGF-beta) superfamily, and is an oocyte secreted paracrine factor essential for mammalian ovarian folliculogenesis (1-2). Mouse GDF-9 is synthesized as a 441aa \nprepropeptide that contains a 29aa signal sequence, a 277aa propeptide, and a 135aa mature chain (SwissProt # Q07105). Residues 340-441 constitute a TGF-beta like domain. In addition, there is one potential site of N-linked glycosylation in the mature chain. Unlike other members of the TGF-beta superfamily, GDF-9 lacks the conserved cysteine residue that is believed to form the sole disulfide linkage between subunits in other family members (3). Mature mouse GDF-9 shares 90% aa sequence identity with mature human GDF-9. The protein is expressed throughout the development of the maturing follicle (2). GDF-9 functions as a paracrine factor in the regulation of granulosa cell proliferation and differentiation, and is essential for fertility (2, 4). Studies on GDF-9 null mice have demonstrated arrested follicular development at the primary follicle stage (5). Mouse GDF-9 induces Smad2 phosphorylation and inhibin production in rat \ndiethylstilbestrol-treated granulosa cells (6) and in human granulosa-luteal cells (7). The downstream signaling actions of GDF-9 are mediated by the type I receptor, activin receptor-like kinase 5 (ALK5), initiating the subsequent activation of Smad2 and Smad3 (2, 8). GDF-9 uses the BMP type II receptor (BMPRII) as its other signaling receptor (2, 9). \n\nSource: \nDNA sequence encoding mouse GDF-9 (Met 1-Arg 441; Accession # AAA53035) was expressed in Chinese Hamster Ovary (CHO) cells. \n \nMolecular Mass: Based on N-terminal amino acid sequencing, the recombinant mouse GDF-9 starts at Gly 307, and has a predicted molecular mass of 15.6kD. The recombinant mouse GDF-9 migrates as an approximately 20kD protein in SDS-PAGE under reducing conditions. \n\n100ng/mL of rmGDF-9 can effectively induce Smad2 phosphorylation.\n\nActivity: \nMeasured by its ability to induce Smad2 phosphorylation in P19 cells (Mazerbourg, S., et al. 2004. Mol. Endocrinol. 18:653). 100ng/ml of rmGDF-9 can effectively induce Smad2 phosphorylation.\n\nEndotoxin: (same/less than) 1.0 EU/ 1ug (LAL). \n\nStorage and Stability:\n12 months from date of receipt, 20 to 70 degrees C as supplied.\n1 month, 2 to 8 degrees C under sterile conditions after reconstitution.\n3 months, 20 to 70 degrees C under sterile conditions after reconstitution.\nUse a manual defrost freezer and avoid repeated freeze-thaw cycles.

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SPECIFICATIONS

Catalog Number

G8989-49E

Size

10ug

Form

Supplied as a lyophilized powder in 0.2um sterile-filtered solution, 4mM HCl, 50ug BSA/1ug of protein. Reconstitute with sterile 4mM HCl, 0.1% BSA.

Purity

Purified (same/more than) 95% as determined by SDS-PAGE under reducing conditions and visualized by silver stain.

References

1. McGrath, S.A. et al., 1995, Mol. Endocrinol. 9:131. 2. Mottershead, D.G. et al., 2008, Mol. Cell. Endocrinol. 283:58. 3. McPherron, A.C. and S.-J. Lee, 1992, J. Biol. Chem. 268:3444. 4. Gilchrist, R.B. et al., 2006, J. Cell. Sci. 119:3811. 5. Dong, J. et al., 1996, Nature 383:531. 6. Roh, J.S. et al., 2003, Endocrinology 144:172. 7. Kaivo-Oja, N. et al., 2003, J. Clin. Endocrinol. Metab. 88:755. 8. Mazerbourg, S. et al., 2004, Mol. Endocrinol. 18:653. 9. Vitt, U.A. et al., 2002, Biol. Reprod. 67:473.