SGK1 antibody | knockout validation | Cell Signaling 12103

This is a knockout-validated antibody summary, based on the publication "Mitochondrial Permeability Uncouples Elevated Autophagy and Lifespan Extension", as cited below [1]. 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: SGK1

Catalog number: 12103

Summary: Rabbit monoclonal IgG against a synthetic peptide corresponding to residues surrounding Gly420 of human SGK1 protein. Reacts with human by western blot and immunoprecipitation.

Validation Method

Western blot

Sample

Control, Sgk1-knockout AML12 hepatocytes and primary hepatocytes from Sgk1 liver-specific knockout. Cell lysates were prepared in RIPA buffer (50 mM Tris-HCl pH 7.4,150 mM NaCl, 1% Triton x-100, 1% Sodium deoxycholate, 0.1% SDS,1 mM EDTA with proteinase inhibitor cocktail from Roche).

Primary incubation

According to manufacturers’ protocols.

Secondary incubation

1:5,000 dilution goat-anti-rabbit-HRP conjugate (GE Healthcare) in 5% BSA dissolved in TBST.

Detection

West-Pico chemiluminescence substrate (Thermo Pierce).

Figure

Western blot of the indicated proteins in Sgk1 knockout (Sgk1(-/-)) AML12 hepatocytes and primary hepatocytes from Sgk1 liver-specific knockout (Sgk1Lko) versus Sgk1flox/flox control mice (control). Please see Figure 1i and 4b in the article [1].

References
  1. Zhou B, Kreuzer J, Kumsta C, Wu L, Kamer K, Cedillo L, et al. Mitochondrial Permeability Uncouples Elevated Autophagy and Lifespan Extension. Cell. 2019;177:299-314.e16 pubmed publisher