Beta catenin antibody | knockout validation | Abcam ab32572

This is a knockout-validated antibody summary, based on the publication "Hepatocyte-Specific β-Catenin Deletion During Severe Liver Injury Provokes Cholangiocytes to Differentiate Into Hepatocytes", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Beta catenin antibody | knockout validation | Abcam ab32572 figure 1
Figure 1. Immunofluorescence of the indicated proteins in both control WT2 and KO2 mice left on normal diet for 6 months (scale bar 50 μm). From [1].
Antibody information

Rabbit monoclonal IgG

Company: Abcam

Antibody: beta Catenin

Catalog number: ab32572

Summary: Rabbit monoclonal IgG against a synthetic peptide within human beta Catenin(aa 1-100, N terminal). Reacts with rat, hamster, human, and African green monkey. Predicted to work with mouse, sheep, cow, macaque monkey. Suitable for western blot, immunoprecipitation (IP), immunohistochemistry (paraffin), immunocytochemistry/immunofluorescence and Chromatin IP. Unsuitable for flow cytometry.

Beta catenin antibody | knockout validation | Abcam ab32572 figure 2
Figure 2. Tiled IHC β-catenin staining of a lobe from a KO2 mouse left on normal diet for 6 months after AAV8 injection reveals virtually no β-catenin-positive hepatocytes. From [1].
Validation Method

Immunocytochemistry | Immunohistochemistry

Sample

WT and beta catenin-KO mouse liver. ICC: Tissue samples were drop fixed in 10% buffered formalin overnight, cryopreserved in 30% sucrose in PBS overnight, frozen in OCT compound. IHC: Tissue samples were drop-fixed in 10% buffered formalin for 48 hours prior to paraffin embedding.

Blocking agent

ICC: 2% Donkey serum in 0.1% Tween™ 20 in PBS (antibody diluent) for 30 minutes at room temperature.

IHC: 3% H2O2 for 10 minutes to quench endogenous peroxidase activity. After washing with PBS, slides were blocked with Super Block (ScyTek Laboratories, AAA500) for 10 minutes.

Primary incubation

ICC: 1:100 dilution at 4℃ overnight.

ICC: 1:100 dilution for one hour at room temperature.

Secondary incubation

ICC: proper fluorescent secondary antibody (AlexaFluor 488/555/647, Invitrogen) diluted 1:800 in antibody diluent for two hours at room temperature.

IHC: appropriate biotinylated secondary antibody (Vector Laboratories) diluted 1:500 in antibody diluent for 30 minutes at room temperature.

Detection

ICC: Nikon Eclipse Ti epifluorescence microscope or a Zeiss LSM700 confocal microscope.

IHC: DAB Peroxidase Substrate Kit (Vector Laboratories, SK-4100) and images

were taken on a Zeiss Axio Observer.Z1 microscope.

Clone note

The same clone (E247) is sold as Abcam ab32572, ab194118, ab194120; Invitrogen MA5-14461; GeneTex GTX61089; EMD Millipore 04-1002.

References
  1. Russell J, Lu W, Okabe H, Abrams M, Oertel M, Poddar M, et al. Hepatocyte-Specific β-Catenin Deletion During Severe Liver Injury Provokes Cholangiocytes to Differentiate Into Hepatocytes. Hepatology. 2019;69:742-759 pubmed publisher