This is a knockout-validated antibody summary, based on the publication "AKT isoform-specific expression and activation across cancer lineages", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

[enlarge]

Figure 1. Western blot of AKT2 and pAKT2 in HCT116 parental, AKT2−/−, and AKT1−/− cells serum starved overnight and treated (or not treated) with insulin (20 μg/ml) for 30 min. ERK immunoblotting showed equivalent loading. From [1].

Rabbit monoclonal IgG

Company: Cell Signaling

Antibody: Phospho-Akt2 (Ser474)

Catalog number: 8599

Summary: Rabbit monoclonal IgG against a synthetic phosphopeptide corresponding to residues surrounding Ser474 of human Akt2 protein. Reacts with human, mouse and rat. Suitable for western blot and immunoprecipitation.

Western blot

HCT116 parental, AKT2−/−, and AKT1−/− cells. Cells were lysed in radioimmunoprecipitation assay (RIPA) buffer (50 mM Tris-HCl (pH 7.4), 150 mM NaCl, 1% NP-40, 0.5% sodium deoxycholate, and 0.1% sodium dodecyl sulfate), supplemented with protease inhibitor and phosphatase inhibitor cocktails (Pierce Biotechnology, Rockford, IL).