Phospho-AKT1 (Thr34) Rabbit Polyclonal Antibody

• Catalog Number: orb6783
• Category: Antibodies
• Description: Phospho-AKT1 (Thr34) Rabbit Polyclonal Antibody
• Target: AKT1
• Clonality: Polyclonal
• Species/Host: Rabbit
• Isotype: IgG
• Conjugation: Unconjugated
• Reactivity: Human, Mouse
• Predicted Reactivity: Rat
• 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 Thr34 DG(p-T)FI
• UniProt ID: P31749
• RRID: AB_10919593
• 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=2μg/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: AKT1(phospho T34); p-AKT1(phospho T34); AKT 1; AKT
• 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 (Thr34) antibody (orb6783, Green), dilution: 3 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.

Blank control: A549. Primary Antibody (green line): Rabbit Anti-phospho-AKT1 (Thr34) antibody (orb6783), 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-AKT1 (Thr34)) Polyclonal Antibody, Unconjugated (orb6783) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Sample: Lane 1: SiHa (Human) Cell Lysate at 30 ug, Lane 2: NIH/3T3 (Mouse) Cell Lysate at 30 ug, Lane 3: Adrenal glands (Mouse) Lysate at 40 ug, Lane 4: Skeletal muscle (Mouse) Lysate at 40 ug, Lane 5: Ovary (Mouse) Lysate at 40 ug, Lane 6: Lung (Mouse) Lysate at 40 ug, Lane 7: Cerebrum (Mouse) Lysate at 40 ug.