ab93892 staining GDF 9 in Rat ovary tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and blocked with 1.5% serum for 1 hour at 24°C; antigen retrieval was enzymatic. Samples were incubated with primary antibody (1μg/ml) for 12 hours at 4°C. A Biotin-conjugated Goat anti-rabbit IgG polyclonal (1/200) was used as the secondary antibody.See Abreview
Anti-GDF 9 antibody (ab93892) at 1 µg/ml + Ovary (Human) Tissue Lysate - adult normal tissue (ab30222) at 10 µgSecondaryGoat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.
IHC image of GDF 9 staining in human normal testis FFPE section, performed on a BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab93892, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX
ICC/IF image of ab93892 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab93892 at 5µg/ml overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti- rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Immunohistochemical analysis of oocytes of primordial (A), and early antral follicles (B) of sheep ovary tissue, staining GDF 9 with ab93892.Tissue was fixed with paraformaldehyde and blocked with 0.4% blocking agent for 5 minutes at 20°C; antigen retrieval was by heat mediation in citrate buffer (pH 6). Samples were incubated with primary antibody (1/1500 in diluent) for 24 hours at 4°C. An undiluted HRP-conjugated goat anti-rabbit polyclonal IgG was used as the secondary antibody.See Abreview