Phospho-AKT1 (Tyr474) Rabbit Polyclonal Antibody

• Catalog Number: orb155627
• Category: Antibodies
• Description: Phospho-AKT1 (Tyr474) Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, ICC, IF, IHC-Fr, IHC-P, WB
• Predicted Reactivity: Bovine, Canine, Gallus, Porcine, Rabbit, Rat, Sheep
• Reactivity: Human, Mouse
• Isotype: IgG
• Immunogen: KLH conjugated synthesised phosphopeptide derived from human AKT1 around the phosphorylation site of Tyr474 FS(p-Y)SA
• Antibody Type: Primary Antibody
• Concentration: 1mg/ml
• Dilution range: WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:100-500, ICC/IF=1:100, IF=1:100-500, Flow-Cyt=1μg/Test
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 56 kDa
• Target: AKT1
• UniProt ID: P31749
• 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: AKT1(phospho Y474); AKT1(phospho-Tyr474); AKT3(pho
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Blank control: A549. Primary Antibody (green line): Rabbit Anti-phospho-AKT1 (Tyr474) antibody (orb155627), 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.

Primary Antibody: Rabbit Anti-phospho-AKT1 (Tyr474) antibody, Supplier catalog number: orb155627 (Green), Isotype Control Antibody: Rabbit IgG (orange), Secondary Antibody: Goat anti-rabbit IgG-FITC, Positive control: Hela Cells (blue).

Sample: Hela (Human) Cell Lysate at 30 ug, Primary: Anti-phospho-AKT1 (Tyr474) (orb155627) 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: Lung (Mouse) Lysate at 40 ug, Primary: Anti-phospho-AKT1 (Tyr474) (orb155627) 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: A549 cell, 4% Paraformaldehyde-fixed, Triton X-100 at room temperature for 20 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Antibody incubation with (phospho-AKT1 (Tyr474)) polyclonal Antibody, Unconjugated (orb155627) 1:100, 90 minutes at 37°C, followed by a FITC conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.

Tissue/Cell: human myometrium tissue, 4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, pH 6.0), 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-AKT1 (Tyr474) Polyclonal Antibody, Unconjugated (orb155627) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.