Human GDF-9 Antibody

  
  • Species Reactivity
    Human
  • Specificity
    Detects human GDF-9 in direct ELISAs.
  • Source
    Monoclonal Mouse IgG2B Clone # 917319
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    Human embryonic kidney cell line derived recombinant human GDF-9
    Met1-Arg454
    Accession # O60383
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Flow Cytometry
    0.25 µg/106 cells
    See below
  • CyTOF-ready
    Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
  • Immunocytochemistry
    8-25 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Detection of GDF‑9 in OVCAR‑3 Human Cell Line by Flow Cytometry. OVCAR‑3 human ovarian carcinoma cell line was stained with Mouse Anti-Human GDF‑9 Monoclonal Antibody (Catalog # MAB8266, filled histogram) or isotype control antibody (Catalog # MAB0041, open histogram), followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005).
Immunocytochemistry
GDF‑9 in OVCAR‑3 Human Cell Line. GDF‑9 was detected in immersion fixed OVCAR‑3 human ovarian carcinoma cell line using Mouse Anti-Human GDF‑9 Monoclonal Antibody (Catalog # MAB8266) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to secreted molecule. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.5 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 12 months from date of receipt, -20 to -70 °C as supplied.
    • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
    • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: GDF-9

Growth Differentiation Factor-9 (GDF-9) is an oocyte secreted paracrine factor in the TGF-beta superfamily (1, 2). It is synthesized as a prepropeptide and is subsequently processed by proteases into the mature protein (1, 2). Mature human GDF-9 has a predicted molecular weight of 16 kDa and shares 89.6% and 91.9% amino acid sequence identity with the mouse and rat orthologs, respectively. Despite the high homology, mouse GDF-9 is secreted in an active form, while human GDF-9 is latent. A single mutation Gly391Arg increases the affinity between human GDF-9 and its signaling receptors and make it more active (3). It forms both non-covalent homodimers and heterodimers with BMP-15, which is coordinately expressed with GDF-9 in the oocyte. (2, 4, 5). GDF-9 signals through TGF-beta RI/ALK-5 and BMPR-II, while the GDF-9:BMP-15 heterodimer is believed to signal through BMPR-II, ALK 4/5/7, and BMPR-IB/ALK-6 (5-8). SMAD2 and SMAD3 are phosphorylated following activation of receptor complexes by GDF-9 (5, 6). GDF-9 functions as a paracrine factor in the development of primary follicles in the ovary. It is critical for the growth of granulosa and theca cells and for the differentiation and maturation of the oocyte (5, 9-11). GDF-9 is thought to act synergistically with BMP-15 to control development of the oocyte-cumulus cell complex (4-6). In humans, GDF-9:BMP-15 heterodimers have been shown to be more potent regulators of granulosa cell functions compared to GDF-9 homodimers (6). Aberrant GDF-9 expression and activation is associated with a multitude of common human ovarian disorders including premature ovarian failure and polycystic ovary syndrome (10, 12-14). In breast and bladder cancers, GDF-9 is believed to function as a tumor suppressor because its expression levels are inversely correlated with the aggressiveness of the cancer (15, 16). In prostate cancer, however, GDF-9 may enhance tumor progression by promoting tumor cell growth and epithelial-to-mesenchymal transition (17, 18).

  • References:
    1. McGrath, S. A. et al. (1995) Mol. Endocrinol. 9:131.
    2. Aaltonen, J. et al. (1999) J. Clin. Endocrinol. Metab. 84:2744.
    3. Simpson, C.M. et al. (2012) 153:1301.
    4. Liao, W.X. et al. (2003) J. Biol. Chem. 278:3713.
    5. Gilchrist, R.B. et al. (2008) Hum. Reprod. Update 14:159.
    6. Peng, J. et al. (2013) Proc. Natl. Acad. Sci. USA 110:E776.
    7. Vitt, U.A. et al. (2002) Biol. Reprod. 67:473.
    8. Mazerbourg, S. et al. (2004) Mol. Endocrinol. 18:653.
    9. Hreinsson, J.G. et al. (2002) J. Clin. Endocrinol. Metab. 87:316.
    10. Otsuka, F. et al. (2011) Mol. Reprod. Dev. 78:9.
    11. Dong, J. et al. (1996) Nature 383:531.
    12. Zhao, S.Y. et al. (2010) Fertil. Steril. 94:261.
    13. Wei, L.N. et al. (2011) Fertil. Steril. 96:464.
    14. Simpson, C.M. et al. (2014) J. Clin. Endocrinol. Metab. [Epub ahead of print].
    15. Hanavadi, S. et al. (2007) Ann. Surg. Oncol. 14:2159.
    16. Du, P. et al. (2012) Int. J. Mol. Med. 29:428.
    17. Bokobza, S.M. et al. (2010) J. Cell. Physiol. 225:529.
    18. Bokobza, S.M. et al. (2011) Mol. Cell. Biochem. 349:33.
  • Long Name:
    Growth Differentiation Factor 9
  • Entrez Gene IDs:
    2661 (Human); 14566 (Mouse)
  • Alternate Names:
    GDF9; GDF-9; growth differentiation factor 9; growth/differentiation factor 9
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