This is a knockout-validated antibody summary, based on the publication "Greatwall-phosphorylated Endosulfine is both an inhibitor and a substrate of PP2A-B55 heterotrimers", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Figure 1. Western blot of extracts from Drosophila larvae mutant for the gene encoding B55 (twins) regulatory subunit of PP2A. Tubulin was used as loading control. From [1].

Rabbit monoclonal

Company: Cell Signaling

Antibody: Twins

Catalog number: 2290

Summary: Rabbit monoclonal raised against a synthetic peptide corresponding to the sequence of human PP2A B subunit. Reacts with human, mouse, rat, monkey, and D. melanogaster. It is predicted to react with chicken, bovine, and pig, based on 100% sequence homology.

Western blot

Drosophila larvae mutant for the gene encoding B55 (twins). Larvae were homogenized in ice-cold phosphatase buffer (50 mM Tris, pH 7.5, 150 mM NaCl, 1 mM EDTA, 0.1 mM EGTA, 0.25% NP-40) to which was added a 1:100 dilution of phenylmethanesulfonyl fluoride (PMSF; Thermo Scientific), a 1:50 dilution of Halt protease inhibitor cocktail (Thermo Scientific), and a 1:100 dilution of a 100 mg/ml stock solution of soybean trypsin inhibitor (AMRESCO, Solon, OH; catalog number M191).

Goat anti-rabbit IgG (H+L)-HRP conjugate (catalog #170-6515; Bio-rad, Hercules, CA) at a 1:5,000 dilution.

ECL Western Blotting Substrate (Thermo Scientific).

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.