Product nameAnti-WAC antibody
See all WAC primary antibodies
DescriptionRabbit polyclonal to WAC
Tested applicationsSuitable for: WB, ICC/IFmore details
Species reactivityReacts with: 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.
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.
Storage instructionsShipped 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.
Storage bufferpH: 7.40
Preservative: 0.02% Sodium azide
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.
Concentration information loading...
PurityImmunogen affinity purified
Our Abpromise guarantee covers the use of ab109486 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 83 kDa (predicted molecular weight: 71 kDa).|
|ICC/IF||Use a concentration of 5 µg/ml.|
FunctionActs 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.
Sequence similaritiesContains 1 WW domain.
modificationsPhosphorylated on tyrosine residues.
Cellular localizationNucleus speckle. Nucleus. In distinct nuclear speckles. Colocalizes with pre-mRNA processing complexes.
- Information by UniProt
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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.
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.
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 71 kDa
Observed band size: 85 kDa why is the actual band size different from the predicted?
Exposure time: 20 minutes
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.
ab109486 has not yet been referenced specifically in any publications.