GDF-9 Antibody (917319) [Unconjugated]

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).

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.

• Reactivity: Hu
• Applications: Flow, CyTOF-ready, ICC/IF
• Clone: 917319
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated

GDF-9 Antibody (917319) [Unconjugated] Summary

• Immunogen: Human embryonic kidney cell line derived recombinant human GDF-9
  Met1-Arg454
  Accession # O60383
• Specificity: Detects human GDF-9 in direct ELISAs.
• Source: N/A
• Isotype: IgG2b
• Clonality: Monoclonal
• Host: Mouse
• Gene: GDF9
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • Flow Cytometry 0.25 ug/10^6 cells
  • Immunocytochemistry 8-25 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for GDF-9 Antibody (917319) [Unconjugated]

• GDF9
• GDF-9
• growth differentiation factor 9
• growth/differentiation factor 9

Background

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).

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

• Gene Symbol: GDF9
• Uniprot:
  • O60383()