Key features and details
- Rabbit monoclonal [EP2297Y] to DAB2
- Suitable for: WB, IP, Flow Cyt, ICC
- Reacts with: Human
- Isotype: IgG
Product nameAnti-DAB2 antibody [EP2297Y]
See all DAB2 primary antibodies
DescriptionRabbit monoclonal [EP2297Y] to DAB2
Tested applicationsSuitable for: WB, IP, Flow Cyt, ICCmore details
Species reactivityReacts with: Human
Synthetic peptide within Human DAB2 aa 650-750 (C terminal). The exact sequence is proprietary.
- WB: PC3 and HeLa whole cell lysates; ICC: HeLa cells; IP: HeLa whole cell lysate; Flow Cyt: HeLa cells.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 0.05% BSA, 40% Glycerol, 59% PBS
Concentration information loading...
PurityProtein A purified
Our Abpromise guarantee covers the use of ab76253 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/1000. Detects a band of approximately 105 kDa (predicted molecular weight: 83 kDa).
For unpurified use at 1/2000 - 1/10000.
For unpurified use at 1/50.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
For unpurified use at 1/100 - 1/250.
RelevanceDAB2 is a component of the CSF1 signal transduction pathway. DAB2 mRNA is expressed in normal ovarian epithelial cells but is down regulated or absent from ovarian carcinoma cell lines. The down regulation of DAB2 may play an important role in ovarian carcinogenesis. This gene was initially named DOC2 (for Differentially expressed in Ovarian Cancer) and is distinct from the DOC2A and DOC2B genes (for double C2 like domains, alpha and beta).
Cellular localizationCytoplasmic vesicle; clathrin coated vesicle; clathrin coated vesicle membrane. Cell membrane; coated pit; clathrin coated pit. Note: Colocalizes with large insert containing isoforms of MYO6 at clathrin coated pits/vesicles.
- DAB 2 antibody
- Dab, mitogen-responsive phosphoprotein, homolog 2 (Drosophila) antibody
- DAB2 antibody
All lanes : Anti-DAB2 antibody [EP2297Y] (ab76253) at 1/1000 dilution (purified)
Lane 1 : PC-3 (Human prostate adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Lysates/proteins at 20 µg per lane.
All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Predicted band size: 83 kDa
Observed band size: 105 kDa why is the actual band size different from the predicted?
Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling DAB2 with purified ab76253 at 1/100 dilution (1.1 µg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% TritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling DAB2 with purified ab76253 at 1/100 dilution (1 µg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor®488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
Purified ab76253 at 1/20 dilution (0.5 µg) immunoprecipitating DAB2 in HeLa whole cell lysate.
Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10 µg
Lane 2 (+): ab76253 + HeLa whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab76253 in HeLa whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: 105 kDa
ab76253 (unpurified) stained HeLa cells. The cells were 4% formaldehyde fixed for 10 minutes at room temperature and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (unpurified ab76253 at 1/100 dilution) overnight at +4°C. The secondary antibody (pseudo-colored green) was Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) used at a 1/1000 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour 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.
Overlay histogram showing HeLa cells stained with unpurified ab76253 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab76253, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
ab76253 has been referenced in 2 publications.
- Sun C et al. miR-106b targets DAB2 to promote hepatocellular carcinoma cell proliferation and metastasis. Oncol Lett 16:3063-3069 (2018). PubMed: 30127897
- Xu S et al. Loss of Dab2 expression in breast cancer cells impairs their ability to deplete TGF-ß and induce Tregs development via TGF-ß. PLoS One 9:e91709 (2014). IHC ; Human . PubMed: 24638085