Western blot analysis of mouse lung tissue using AKT (phospho-Thr308) antibody.
Western blot analysis of extracts from Heart,using phospho-Akt(thr308) antibody..
Blank control (blue line): A431 (Black). Primary Antibody (green line): Rabbit Anti-phospho-Akt (Thr308) antibody (orb14786), Dilution: 1 µg/10 6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody (white blue line): Goat anti-rabbit IgG-PE, Dilution: 1 µg/Test. Protocol, The cells were fixed with 4% paraformaldehyde (10 min), then permeabilized with 90% ice-cold methanol for 20 min on ice. Cells stained with Primary Antibody for 30 min at room temperature. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions followed by the antibody for 15 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20000 events was performed.
Paraformaldehyde-fixed, paraffin embedded (Mouse brain), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37C for 30 min, Antibody incubation with (phospho-Akt (Thr308)) Polyclonal Antibody, Unconjugated (orb14786) at 1:400 overnight at 4C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
Paraformaldehyde-fixed, paraffin embedded (Rat brain), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37C for 30 min, Antibody incubation with (phospho-Akt (Thr308)) Polyclonal Antibody, Unconjugated (orb14786) at 1:400 overnight at 4C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
Sample: Liver (Mouse) Lysate at 30 ug, Primary: Anti-phospho-Akt (Thr308) (orb14786) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-Mouse IgG at 1/10000 dilution, Predicted band size: 56 kD, Observed band size: 56 kD.
Western blot analysis of extracts from Heart, using phospho-Akt (thr308) Antibody.
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