Detection of Mouse and Hamster GDF‑9 by Western Blot. Western blot shows lysates of human ovary tissue and CHO Chinese hamster ovary cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Mouse/Hamster GDF‑9 Antigen Affinity-purified Polyclonal Antibody |
(Catalog # AF739) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for GDF‑9 at approximately 18 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
|GDF‑9 in Mouse Ovary. GDF‑9 was detected in perfusion fixed frozen sections of mouse ovary using Goat Anti-Mouse/Hamster GDF‑9 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF739) at 5 µg/mL overnight at 4 °C. Tissue was stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to developing oocytes. View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.|
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 441 amino acid (aa) prepropeptide that contains a 29 aa signal sequence, a 277 aa propeptide, and a 135 aa mature chain. 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 diethylstilbestrol 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).
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