Phospho-AKT1 + AKT2 + AKT3 (Thr308+Thr309+Thr305) Rabbit Polyclonal Antibody

• Catalog Number: orb6780
• Category: Antibodies
• Description: Phospho-AKT1 + AKT2 + AKT3 (Thr308+Thr309+Thr305) 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, Sheep
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: KLH conjugated Synthesised phosphopeptide derived from human AKT1 around the phosphorylation site of Thr308 MK(p-T)FC
• 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: 53 kDa
• Target: AKT1
• UniProt ID: P31749
• RRID: AB_10931973
• 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 T308); AKT2(phospho T309); AKT3(phosp
• 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 (Thr308+Thr309+Thr305) antibody (orb6780, 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 (Thr308+Thr309+Thr305)) Polyclonal Antibody, Unconjugated (orb6780) at 1:400 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-AKT1 + AKT2 + AKT3 (Thr308+Thr309+Thr305)) Polyclonal Antibody, Unconjugated (orb6780) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.Antibody, Unconjugated (orb6780) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.

Sample: Hela (Human) Cell Lysate at 30 ug, HepG2 (Human) Cell Lysate at 30 ug, Primary: Anti-phospho-AKT1 + AKT2 + AKT3 (Thr308+Thr309+Thr305) (orb6780) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 62/53 kD, Observed band size: 62/53 kD.

Sample: Lung (Mouse) Lysate at 40 ug, Primary: Anti-phospho-AKT1 + AKT2 + AKT3 (Thr308+Thr309+Thr305) (orb6780) at 1/300 dilution, Secondary: HRP conjugated Goat-Anti-rabbit IgG (orb572747) at 1/5000 dilution, Predicted band size: 53 kD, Observed band size: 53 kD.

Sample: NIH/3T3 (Mouse) Cell Lysate at 30 ug, Adrenal gland (Mouse) Lysate at 40 ug, MCF-7 (Human) Cell Lysate at 30 ug, A549 (Human) Cell Lysate at 30 ug, Primary: Anti-phospho-AKT1+AKT2+AKT3 (Thr308+Thr309+Thr305) (orb6780) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 60 kD, Observed band size: 60 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 + AKT2 + AKT3 (Thr308+Thr309+Thr305)) polyclonal Antibody, Unconjugated (orb6780) 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.