Phospho-AKT (Ser473) Rabbit Polyclonal Antibody

• Catalog Number: orb11293
• Category: Antibodies
• Description: Phospho-AKT (Ser473) Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, IF, IHC-Fr, IHC-P, WB
• Predicted Reactivity: Bovine, Canine, Gallus, Porcine, Rabbit, Sheep
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: KLH conjugated Synthesised phosphopeptide derived from human AKT around the phosphorylation site of Ser473 QF(p-S)YS
• Concentration: 1mg/ml
• Dilution range: WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:100-500, IF=1:100-500, Flow-Cyt=2ug/Test
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 56 kDa
• Target: AKT1
• UniProt ID: P31749
• RRID: AB_10752490
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Alternative names: Akt(Phospho-Ser473); AKT (phospho-S473); AKT (phos
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Blank control: A549. Primary Antibody (green line): Rabbit Anti-phospho-AKT (Ser473) antibody (orb11293), Dilution: 2 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody: Goat anti-rabbit IgG-FITC, Dilution: 1 µg/Test. Protocol, The cells were incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20000 events was performed.

Blank control: A549. Primary Antibody (green line): Rabbit Anti-phospho-AKT (Ser473) antibody (orb11293), Dilution: 2 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody: Goat anti-rabbit IgG-FITC, Dilution: 1 µg/Test. Protocol, The cells were incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 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 37°C for 30 min, Antibody incubation with (phospho-AKT (Ser473)) Polyclonal Antibody, Unconjugated (orb11293) at 1:2000 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

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 37°C for 30 min, Antibody incubation with (phospho-AKT (Ser473)) Polyclonal Antibody, Unconjugated (orb11293) at 1:2000 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand 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 37°C for 30 min, Antibody incubation with (phospho-AKT (Ser473)) Polyclonal Antibody, Unconjugated (orb11293) at 1:2000 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand 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 37°C for 30 min, Antibody incubation with (phospho-AKT (Ser473)) Polyclonal Antibody, Unconjugated (orb11293) at 1:2000 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (rat lung), 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 37°C for 30 min, Antibody incubation with (phospho-AKT (Ser473)) Polyclonal Antibody, Unconjugated (orb11293) at 1:2000 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Sample: Heart (Mouse) Lysate at 40 ug, Primary: Anti-phospho-AKT (Ser473) (orb11293) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 56 kD, Observed band size: 56 kD.

Sample: Liver (Mouse) Lysate at 30 ug, Primary: Anti-phospho-AKT (Ser473) (orb11293) at 1:300 dilution, Secondary: HRP conjugated Goat Anti-Rabbit IgG (orb572747) at 1:5000 dilution, Predicted band size: 56kD, Observed band size: 56kD.

Sample: U937 Cell (Human) Lysate at 30 ug, Primary: Anti-phospho-AKT (Ser473) (orb11293) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 56 kD, Observed band size: 56 kD.

Tissue/Cell: Human breast, 4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min, Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Incubation: Anti-phospho-AKT/PKB (Ser473) Polyclonal Antibody, Unconjugated (orb11293) 1:600, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.