Human, Zebrafish Predicted to work with:
Sheep, Chicken, Cow, Cat, Pig, Monkey
Synthetic peptide corresponding to Human Inhibin beta B aa 350 to the C-terminus conjugated to Keyhole Limpet Haemocyanin (KLH). Database link: P09529 (Peptide available as ab91445)
This antibody gave a postive signal in the following tissue lysates: Human testis; Human pituitary gland; Human adipose tissue. It also gave a positive signal in human testis tissue sections.
This antibody gave a positive result in IF in the following formaldehyde fixed cell lines: DU145.
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Use a concentration of 5 µg/ml.
Use a concentration of 1 µg/ml. Detects a band of approximately 52 kDa (predicted molecular weight: 45 kDa).
Inhibins and activins inhibit and activate, respectively, the secretion of follitropin by the pituitary gland. Inhibins/activins are involved in regulating a number of diverse functions such as hypothalamic and pituitary hormone secretion, gonadal hormone secretion, germ cell development and maturation, erythroid differentiation, insulin secretion, nerve cell survival, embryonic axial development or bone growth, depending on their subunit composition. Inhibins appear to oppose the functions of activins.
All lanes : Anti-Inhibin beta B antibody (ab69286) at 1 µg/ml
Lane 1 : Human testis tissue lysate - total protein (ab30257) Lane 2 : Brain: pituitary gland (Human) Whole Cell Lysate - adult normal tissue (ab29749) Lane 3 : Human adipose normal tissue lysate - total protein (ab28980)
Lysates/proteins at 10 µg per lane.
Secondary Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution Developed using the ECL technique
Western blot - Active human Activin B full length protein (ab50052)
Anti-Inhibin beta B antibody (ab69286) at 1 µg/ml + Recombinant human Activin B protein (ab50052) at 0.001 µg
Secondary Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution Developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 45 kDa
Exposure time : 20 seconds
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Inhibin beta B antibody (ab69286)
IHC image of ab69286 staining in human testis formalin fixed paraffin embedded tissue section, performed on a Leica 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 ab69286, 5µg/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.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Immunocytochemistry/ Immunofluorescence - Anti-Inhibin beta B antibody (ab69286)
ICC/IF image of ab69286 stained DU145 cells. The cells were 4% formaldehyde fixed (5 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 ab69286 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) 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.
Western blot - Anti-Inhibin beta B antibody (ab69286)
All lanes : Anti-Inhibin beta B antibody (ab69286) at 2 µg/ml
Lane 1 : Marker Lane 2 : Zebrafish brain homogenate at 20 µg Lane 3 : Zebrafish heart homogenate at 20 µg
Secondary Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/10000 dilution Developed using the ECL technique