β-Catenin antibody | knockout validation | Cell Signaling 8480

This is a knockout-validated antibody summary, based on publications "SIRT2 deacetylase regulates the activity of GSK3 isoforms independent of inhibitory phosphorylation" [1] (see figure 3a in the article) and "One-step generation of conditional and reversible gene knockouts" [2] (discussed below). Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Antibody information

Rabbit monoclonal IgG

Company: Cell Signaling

Antibody: β-Catenin

Catalog number: 8480

Summary: Rabbit monoclonal IgG against a synthetic peptide corresponding to residues surrounding Pro714 of human β-catenin protein. Reacts with human, mouse, rat, and monkey. It is predicted to react based on 100% sequence homology with zebrafish, bovine, pig, guinea pig, and horse. Suitable for western blot, immunoprecipitation (IP), immunohistochemistry (paraffin and frozen), immunofluorescence/immunocytochemistry, flow cytometry and chromatin IP.

Validation Method

Western blot

Sample

Control and β-Catenin-KO mouse embryonic stem cells (mESC) and HEK 293 cell lines. Cells were lysed in buffer containing complete protease-inhibitor cocktail tablets (11697498001, Roche).

Primary incubation

1:1,000 dilution.

Secondary incubation

1:5,000 dilution goat anti-rabbit HRP conjugated (Cell Signaling).

Detection

ECL prime western blotting detection system (RPN2133, GE Healthcare).

h: Figure

Detection of β-catenin protein by western blotting in control, Cre induction and Cre and Flp dual induction in mESCs and HEK 293 cells. Please see Supplementary Figures 4d and 11g in the article [2].

References
  1. Sarikhani M, Mishra S, Maity S, Kotyada C, Wolfgeher D, Gupta M, et al. SIRT2 deacetylase regulates the activity of GSK3 isoforms independent of inhibitory phosphorylation. elife. 2018;7: pubmed publisher
  2. Andersson Rolf A, Mustata R, Merenda A, Kim J, Perera S, Grego T, et al. One-step generation of conditional and reversible gene knockouts. Nat Methods. 2017;14:287-289 pubmed publisher