Mouse, Human Predicted to work with:
Rat, Rabbit, Horse, Cow, Chimpanzee, Macaque monkey, Gorilla, Chinese hamster, Orangutan
Synthetic peptide corresponding to Human WAC aa 450-550 conjugated to Keyhole Limpet Haemocyanin (KLH). Database link: Q9BTA9
This antibody gave a positive signal in HeLa Nuclear lysate as well as the following whole cell lysates: HeLa; HEK293; NIH3T3.
This antibody gave a positive result when used in the following methanol fixed cell lines: MCF-7.
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.
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 83 kDa (predicted molecular weight: 71 kDa).
Use a concentration of 5 µg/ml.
Acts as a linker between gene transcription and histone H2B monoubiquitination at 'Lys-120' (H2BK120ub1). Interacts with the RNA polymerase II transcriptional machinery via its WW domain and with RNF20-RNF40 via its coiled coil region, thereby linking and regulating H2BK120ub1 and gene transcription. Regulates the cell-cycle checkpoint activation in response to DNA damage.
Contains 1 WW domain.
Phosphorylated on tyrosine residues.
Nucleus speckle. Nucleus. In distinct nuclear speckles. Colocalizes with pre-mRNA processing complexes.
ICC/IF image of ab109486 stained MCF-7 cells. The cells were 100% methanol 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 ab109486 at 5µg/ml overnight at +4°C. The secondary antibody (pseudo-colored green) was Alexa Fluor® 488 goat anti- rabbit (ab150081) IgG (H+L) preadsorbed, used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1h at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.
Western blot - Anti-WAC antibody (ab109486)
All lanes : Anti-WAC antibody (ab109486) at 1 µg/ml (Milk blocking - 3%)
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate Lane 2 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate Lane 3 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate Lane 4 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
Lysates/proteins at 25 µg per lane.
Secondary All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
The predicted molecular weight of WAC is71 kDa (SwissProt), however we expect to observe a banding pattern around 85 kDa. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.
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 3% Milk before being incubated with ab109486 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.