This antibody gave a positive signal in Mouse Lung and Heart tissue lysates, as well as T47D and PANC-1 whole cell lysates.
This antibody gave a positive result when used in the following formaldehyde + methanol fixed cell lines: T47D.
This antibody gave a positive result in IHC in the following FFPE tissue: Human breast adenocarcinoma.
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pH: 7.40 Preservative: 0.02% Sodium azide Constituent: PBS Note: Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml. Detects a band of approximately 43 kDa (predicted molecular weight: 40 kDa).
Use a concentration of 1 µg/ml.
Ligand for members of the frizzled family of seven transmembrane receptors. Probable developmental protein. May be a signaling molecule which affects the development of discrete regions of tissues. Is likely to signal over only few cell diameters.
Belongs to the Wnt family.
Secreted > extracellular space > extracellular matrix.
Wnt9a is expressed in gastric cancer cell lines. The protein encoding this gene shows 75% amino acid identity to chicken Wnt14, which has been shown to play a central role in initiating synovial joint formation in the chick limb. This gene is clustered with another family member, WNT3A, in the chromosome 1q42 region.
Protein Wnt-14 antibody
Protein Wnt-9a antibody
Wingless related MMTV integration site 9A antibody
Wingless type MMTV integration site 9A antibody
Wingless type MMTV integration site family, member 9A antibody
wingless-type MMTV integration site family, member 14 antibody
Wnt 14 antibody
Wnt 9a antibody
Western blot - Anti-Wnt9a antibody (ab125957)
All lanes : Anti-Wnt9a antibody (ab125957) at 1 µg/ml
Lane 1 :Mouse lung normal tissue lysate - total protein (ab29297) Lane 2 : Heart (Mouse) Tissue Lysate Lane 3 :T47D whole cell lysate (ab14899) Lane 4 : PANC-1 (Human Pancreatic Carcinoma) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
Wnt9a contains a potential glycosylation site (SwissProt) which may explain its migration at a higher molecular weight than predicted.
This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab125957 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.
ab125957 stained T47D 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 ab125957 at 1µg/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. This antibody also gave a positive result in 100% methanol fixed (5 min) T47D cells at 1ug/ml.
IHC image of Wnt9a staining in Human breast adenocarcinoma 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 ab125957, 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.
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.