Human/Mouse/Hamster GDF-9 Antibody

Detection of Human/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-Human/Mouse/Hamster GDF-9 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF739) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (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-Human/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; NL001) and counterstained with DAPI (blue). Specific staining was localized to developing oocytes. View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.

Detection of Mouse GDF‑9 by Western Blot. Western Blot shows lysates of mouse ovary. PVDF membrane was probed with 1 µg/ml of Goat Anti-Human/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 17 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Detection of Human GDF-9 by Immunohistochemistry-Paraffin GDF9 is expressed in the human fetal ovary.qRT-PCR analysis of GDF9 (A), BMP15 (B) and NOBOX (C) mRNA expression in human fetal ovary across the gestational range of 8 to 20 weeks. Ovaries (n = 5–7 per group) were grouped according to developmental stage and transcript levels measured relative to those of RPL32. Bars indicate mean±sem. Statistically different levels are indicated by asterisks above the columns, thus expression of GDF9 at 18–20 weeks was significantly higher than at 8–11 weeks (p<0.005) as was expression of BMP15 and of NOBOX (both p<0.01). DAB immunohistochemical detection of GDF9: 19 week (D, E) and 20 week (F) human fetal ovary stained with anti-GDF9 antibody or normal goat IgG negative control (F inset)—positive staining is brown. Thick arrows indicate primordial follicles and thin arrows germ cells that are not stained for GDF9 while the arrowheads indicate primordial follicles that are positive for GDF9. Scale bars are 50μm (D and F) and 20μm (E). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/25790371), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human GDF-9 by Immunocytochemistry/ Immunofluorescence Co-localisation of GDF9 with activin beta A but not DAZL or BOLL prior to follicle formation.(A) Double immunohistochemistry of 18 week fetal ovary stained for GDF9 (green) and activin beta A (red), thus in the merged image co-expression is yellow. Unstained germ cells are indicated with arrows. Counterstain is TOPRO. (B) Triple fluorescent immunohistochemistry for GDF9 (green), DAZL (blue) and BOLL (red) in 20 week human fetal ovary with DAPI as counterstain (grey). Split channel and merged images in (A) and (B) are shown as are merged images of non-immune serum negative control (NEG). Scale bars are 20μm. (C) Nuclear diameters of DAZL, BOLL and GDF9 stained germ cells indicates that GDF9 positive cells are significantly larger (p<0.001) than DAZL but not BOLL expressing cells (bars indicate mean ± sem). (D) Higher magnification merged image of GDF9/DAZL/BOLL immunohistochemistry showing one large primordial follicle is positive for both GDF9 and DAZL but other follicles are positive only for DAZL. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/25790371), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human GDF-9 by Immunohistochemistry-Paraffin GDF9 is expressed in the human fetal ovary.qRT-PCR analysis of GDF9 (A), BMP15 (B) and NOBOX (C) mRNA expression in human fetal ovary across the gestational range of 8 to 20 weeks. Ovaries (n = 5–7 per group) were grouped according to developmental stage and transcript levels measured relative to those of RPL32. Bars indicate mean±sem. Statistically different levels are indicated by asterisks above the columns, thus expression of GDF9 at 18–20 weeks was significantly higher than at 8–11 weeks (p<0.005) as was expression of BMP15 and of NOBOX (both p<0.01). DAB immunohistochemical detection of GDF9: 19 week (D, E) and 20 week (F) human fetal ovary stained with anti-GDF9 antibody or normal goat IgG negative control (F inset)—positive staining is brown. Thick arrows indicate primordial follicles and thin arrows germ cells that are not stained for GDF9 while the arrowheads indicate primordial follicles that are positive for GDF9. Scale bars are 50μm (D and F) and 20μm (E). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/25790371), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human GDF-9 by Immunocytochemistry/ Immunofluorescence Co-localisation of GDF9 with activin beta A but not DAZL or BOLL prior to follicle formation.(A) Double immunohistochemistry of 18 week fetal ovary stained for GDF9 (green) and activin beta A (red), thus in the merged image co-expression is yellow. Unstained germ cells are indicated with arrows. Counterstain is TOPRO. (B) Triple fluorescent immunohistochemistry for GDF9 (green), DAZL (blue) and BOLL (red) in 20 week human fetal ovary with DAPI as counterstain (grey). Split channel and merged images in (A) and (B) are shown as are merged images of non-immune serum negative control (NEG). Scale bars are 20μm. (C) Nuclear diameters of DAZL, BOLL and GDF9 stained germ cells indicates that GDF9 positive cells are significantly larger (p<0.001) than DAZL but not BOLL expressing cells (bars indicate mean ± sem). (D) Higher magnification merged image of GDF9/DAZL/BOLL immunohistochemistry showing one large primordial follicle is positive for both GDF9 and DAZL but other follicles are positive only for DAZL. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/25790371), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human GDF-9 by Immunohistochemistry GDF9 is expressed in the human fetal ovary.qRT-PCR analysis of GDF9 (A), BMP15 (B) and NOBOX (C) mRNA expression in human fetal ovary across the gestational range of 8 to 20 weeks. Ovaries (n = 5–7 per group) were grouped according to developmental stage and transcript levels measured relative to those of RPL32. Bars indicate mean±sem. Statistically different levels are indicated by asterisks above the columns, thus expression of GDF9 at 18–20 weeks was significantly higher than at 8–11 weeks (p<0.005) as was expression of BMP15 and of NOBOX (both p<0.01). DAB immunohistochemical detection of GDF9: 19 week (D, E) and 20 week (F) human fetal ovary stained with anti-GDF9 antibody or normal goat IgG negative control (F inset)—positive staining is brown. Thick arrows indicate primordial follicles and thin arrows germ cells that are not stained for GDF9 while the arrowheads indicate primordial follicles that are positive for GDF9. Scale bars are 50μm (D and F) and 20μm (E). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/25790371), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human GDF-9 by Immunohistochemistry-Paraffin GDF9 is expressed in the human fetal ovary.qRT-PCR analysis of GDF9 (A), BMP15 (B) and NOBOX (C) mRNA expression in human fetal ovary across the gestational range of 8 to 20 weeks. Ovaries (n = 5–7 per group) were grouped according to developmental stage and transcript levels measured relative to those of RPL32. Bars indicate mean±sem. Statistically different levels are indicated by asterisks above the columns, thus expression of GDF9 at 18–20 weeks was significantly higher than at 8–11 weeks (p<0.005) as was expression of BMP15 and of NOBOX (both p<0.01). DAB immunohistochemical detection of GDF9: 19 week (D, E) and 20 week (F) human fetal ovary stained with anti-GDF9 antibody or normal goat IgG negative control (F inset)—positive staining is brown. Thick arrows indicate primordial follicles and thin arrows germ cells that are not stained for GDF9 while the arrowheads indicate primordial follicles that are positive for GDF9. Scale bars are 50μm (D and F) and 20μm (E). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/25790371), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human GDF-9 by Immunohistochemistry-Paraffin GDF9 is expressed in the human fetal ovary.qRT-PCR analysis of GDF9 (A), BMP15 (B) and NOBOX (C) mRNA expression in human fetal ovary across the gestational range of 8 to 20 weeks. Ovaries (n = 5–7 per group) were grouped according to developmental stage and transcript levels measured relative to those of RPL32. Bars indicate mean±sem. Statistically different levels are indicated by asterisks above the columns, thus expression of GDF9 at 18–20 weeks was significantly higher than at 8–11 weeks (p<0.005) as was expression of BMP15 and of NOBOX (both p<0.01). DAB immunohistochemical detection of GDF9: 19 week (D, E) and 20 week (F) human fetal ovary stained with anti-GDF9 antibody or normal goat IgG negative control (F inset)—positive staining is brown. Thick arrows indicate primordial follicles and thin arrows germ cells that are not stained for GDF9 while the arrowheads indicate primordial follicles that are positive for GDF9. Scale bars are 50μm (D and F) and 20μm (E). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/25790371), licensed under a CC-BY license. Not internally tested by R&D Systems.