GDF9 is plays a vital role in ovarian folliculogenesis, follicle development and fertility. Clone 53/1 can be used in assays to detect oocyte expression and has been shown to neutralize GDF9 biological activity. (Gilchrist, R.B. et al.)
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Form in stock: IgG, purified – 1.0 mg/mL. Also available as unpurified supernatant.
Specificity: Tuberculin coupled peptide with sequence VPAKYSPLSVLTIEPDGSIAYKEYEDMIATKC that recognizes an epitope with the EPDG sequence near the C-terminal region of human GDF9.
Human Histology positive control: Ovary
Fusion partner: Spleen cells from immunised Balb/c mice were fused with cells of the SP2/0 myeloma cell line.
Storage: Store at +4°C or -20°C. Avoid repeated freezing and thawing.
Shelf life: 18 months from date of dispatch.
Regulatory/ Restrictions: For research and commercial purposes.
- ⇓ R.B. Gilchrist, L.J. Ritter, M. Cranfield, L.A. Jeffery, F. Amato, S.J. Scott, S. Myllymaa, N. Kaivo-Oja, H. Lankinen, D.G. Mottershead, N.P. Groome, O. Ritvos; Immunoneutralization of Growth Differentiation Factor 9 Reveals It Partially Accounts for Mouse Oocyte Mitogenic Activity. Biol Reprod 2004; 71 (3): 732-739.
- ⇓ Courtney M. Simpson, David M. Robertson, Sara L. Al-Musawi, Derek A. Heath, Kenneth P. McNatty, Lesley J. Ritter, David G. Mottershead, Robert B. Gilchrist, Craig A. Harrison, Peter G. Stanton; Aberrant GDF9 Expression and Activation Are Associated With Common Human Ovarian Disorders. J Clin Endocrinol Metab 2014; 99 (4): E615-E624.
- ⇓ Courtney M. Simpson, Peter G. Stanton, Kelly L. Walton, Karen L. Chan, Lesley J. Ritter, Robert B. Gilchrist, Craig A. Harrison; Activation of Latent Human GDF9 by a Single Residue Change (Gly391Arg) in the Mature Domain. Endocrinology 2012; 153 (3): 1301-1310. WB, Dilution used 1:5000
- ⇓ Li, J.-J., Sugimura, S., Mueller, T. D., White, M. A., Martin, G. A., Ritter, L. J., … Mottershead, D. G. (2015). Modifications of Human Growth Differentiation Factor 9 to Improve the Generation of Embryos From Low Competence Oocytes. Molecular Endocrinology, 29(1), 40–52. WB, Dilution used 1:5000
- ⇓ David G. Mottershead, Minna M. Pulkki, Pranuthi Muggalla, Arja Pasternack, Minna Tolonen, Samu Myllymaa, Olexandr Korchynskyi, Yoshihiro Nishi, Toshihiko Yanase, Stan Lun, Jennifer L. Juengel, Mika Laitinen, Olli Ritvos, Characterization of recombinant human growth differentiation factor-9 signaling in ovarian granulosa cells, Molecular and Cellular Endocrinology, Volume 283, Issues 1–2, 13 February 2008, Pages 58-67, ISSN 0303-7207. WB, Dilution used 1:10000
Clone 53/1 used to detect GDF9 expression in oocyte-medium via western blot
Image caption: GDF9 immunoblot analysis. A) Antibody specificity for GDF9 and (B) detection of oocyte-secreted GDF9. The recombinant proteins indicated (mGDF9 = mouse GDF9, oBMP15 = ovine BMP15) and oocyte extract or oocyte-conditioned medium were subjected to SDS-PAGE immunoblotting with the anti-GDF9 monoclonal antibody and detected using either ECL (A; recombinant proteins) or ECL Advance (B; oocyte products). The 57-kDa band is the GDF9 proprotein while the 17.5-kDa band represents the mature GDF9 monomer. 293H = control; conditioned medium from the untransfected human embryonic kidney 293H parent cell line (Gilchrist, R.B. et al.)
Biological neutralizing capacity of 53/1 by ELISA
Image caption: Biological neutralizing specificity of anti-GDF9-53 with some members of the TGFβ superfamily. Mouse MGC were cultured with human TGFβ1 (0.5 ng/ml), human activin A (50 ng/ml), or mouse GDF9 (40 ng/ml), either in the absence or presence of a high neutralizing dose of 40 μg/ml of mAb-GDF9-53 or 40 μg/ml human IgG. (Gilchrist, R.B. et al.)
Clone 53/1 used to detect GDF9 expression by ELISA
Image caption: GDF9 ELISA. A GDF9 ELISA was developed to measure the amount of GDF9 in HEK-293T conditioned medium. Recombinant mouse GDF9 (●) was used as a standard, and the specificity of the assay was assessed using a range of TGF-β family members; wild-type human GDF9 (♦), human GDF9 L40V (◊), human BMP15 (□), human activin A (○), and human TGF-β3 (▾). Dilutions of concentrated media from cells transfected with empty vector, pcDNA3.1 (r), were included as controls. The ELISA has a specificity of less than 0.1%, with a sensitivity of 0.2 ng/mL. Values represent mean ± SEM in duplicate, from a representative experiment. (Simpson, C et al.)
Clone 53/1 used to detect GDF9 expression in oocyte-medium by western blot
Image caption: … Samples were detected under reducing conditions using GDF9 mAb53 specific for the mature domain. The 60-kDa GDF9 precursor and 20-kDa mature monomer are shown… (Simpson, C et al.)
Dilution used: 1:5000