This is a knockout-validated antibody summary, based on the publication "Hormonal induction and roles of Disabled-2 in lactation and involution", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).
Mouse monoclonal IgG1
Company: BD Transduction Laboratories
Antibody: Dab2
Catalog number: 610465
Summary: Mouse monoclonal IgG1 raised against mouse p96 aa. 31-45. Reacts with mouse, human, and rat. Recommended for western blot, immunofluorescence, and immunohistochemistry. Not recommended for immunoprecipitation.
Western blot | Immunohistochemistry | Immunocytochemistry
WB: Mammary epithelial cells were lysed in a 6-well dish in 0.5 ml of NP-40 IP buffer containing protease inhibitors and phosphatase inhibitors.
IHC/IC: Tissues were fixed with neutral buffered 10% formalin, paraffin embedded, and sectioned on a microtome at 5 µm thickness. Slides were deparaffinized in a graded ethanol series, washed in water and boiled in antigen retrieval solution (10 mM sodium citrate, pH 6.0) prior to staining.
WB: 5% non-fat milk for 1 hour
WB: 1/5,000 overnight
IC: 1/1,000
IHC: 1/1,000
WB: horseradish peroxidase (HRP) at 1/5,000 dilution (mouse secondary antibodies from BioRad; goat secondary antibodies from Jackson ImmunoResearch).
IHC: secondary antibodies HRP-conjugated (Vector Laboratories, CA, USA)
IC: Species-specific secondary antibodies conjugated with Alexa fluorochrome
WB: chemoluminescence detection (SuperSignal West Extended Duration Substrate, Pierce Biotech)
IHC: DAB Peroxidase Substrate Kit (Vector Laboratories, CA, USA) followed by a hematoxylin counterstain.
IC: inverted Zeiss LSM510/uv Axiovert 200 M, laser scanning confocal microscope operated by Zeiss LSM software.
If the antibody described in this summary is a polyclonal antibody, since polyclonal antibodies are of limited quantity, please inquire the supplier whether any current polyclonal antibody with the same catalog number is exactly the same as the one described in this summary. Sometimes, different bleeds or different animals are used, usually with a different lot number. In such cases, the result in this summary may not apply to the new antibody with the same catalog number.