Human GDF-9 Antibody

Catalog # Availability Size / Price Qty
MAB8266
MAB8266-SP
Detection of GDF‑9 in OVCAR‑3  Human Cell Line by Flow Cytometry.
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Human GDF-9 Antibody Summary

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

Flow Cytometry Detection of GDF‑9 in OVCAR‑3  Human Cell Line by Flow Cytometry. View Larger

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. View Larger

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.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
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

Product Datasheets

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Cell and Tissue Staining Kits

Isotype Controls

Reconstitution Buffers

Secondary Antibodies

Mouse IgG2B Antibody

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