Phospho-AKT1+AKT2+AKT3 (Tyr315+316+312) Rabbit Polyclonal Antibody

• Catalog Number: orb6789
• Category: Antibodies
• Description: Phospho-AKT1+AKT2+AKT3 (Tyr315+316+312) Rabbit Polyclonal Antibody
• Target: AKT1
• Clonality: Polyclonal
• Species/Host: Rabbit
• Isotype: IgG
• Conjugation: Unconjugated
• Reactivity: Human, Mouse, Rat
• Predicted Reactivity: Bovine, Canine, Gallus, Porcine, Rabbit, Sheep
• Form/Appearance: Liquid
• Concentration: 1mg/ml
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Purification: Affinity purified by Protein A
• Immunogen: KLH conjugated Synthesised phosphopeptide derived from human AKT1 around the phosphorylation site of Tyr315 PE(p-Y)LA
• UniProt ID: P31749
• RRID: AB_10931712
• MW: 56 kDa
• Tested applications: FC, IF, IHC-Fr, IHC-P, WB
• Dilution range: WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:100-500, IF=1:100-500, Flow-Cyt=1μg/Test
• Antibody Type: Primary Antibody
• 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.
• Alternative names: phospho-AKT1/AKT2/AKT3(Tyr315+Tyr316+Tyr312); AKT1
• Research Area: Akt/PKB Signaling Pathway, Apoptosis, Cell Cycle,
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Blank control (blue): EC9706 (fixed with 2% paraformaldehyde for 10 min at 37°C). Primary Antibody: Rabbit Anti-phospho-AKT1+AKT2+AKT3 (Tyr315+316+312) antibody (orb6789, Green), dilution: 1 µg in 100 µl 1X PBS containing 0.5% BSA, Isotype Control Antibody: Rabbit IgG (orange), used under the same conditions, Secondary Antibody: Goat anti-rabbit IgG-FITC (white blue), dilution: 1:200 in 1X PBS containing 0.5% BSA.

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-AKT1+AKT2+AKT3 (Tyr315+316+312)) Polyclonal Antibody, Unconjugated (orb6789) at 1:500 overnight at 4°C, followed by a conjugated secondary for 20 minutes and DAB staining.

Sample: Heart (Mouse) Lysate at 40 ug, Primary: Anti-phospho-AKT1+AKT2+AKT3 (Tyr315+316+312) (orb6789) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 56 kD, Observed band size: 63 kD.

Sample: Lane 1: Adrenal glands (Mouse) Lysate at 40 ug, Lane 2: Cerebrum (Mouse) Lysate at 40 ug, Primary: Anti-phospho-AKT1+AKT2+AKT3 (Tyr315+316+312) (orb6789) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 60 kD, Observed band size: 60 kD.

Sample: Lane 1: Rat Cerebrum tissue lysates, Lane 2: Rat Heart tissue lysates, Primary: Anti-phospho-AKT1+AKT2+AKT3 (Tyr315+316+312) (orb6789) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 56 kDa, Observed band size: 65 kDa.