AKT ISOFORM antibody

• Catalog Number: orb345936
• Category: Assays and Kits
• Description: AKT ISOFORM antibody
• Species/Host: Mouse
• Clonality: Monoclonal
• Tested applications: ELISA, FC, IHC, WB
• Reactivity: Human
• Isotype: IgG2a
• Immunogen: All Anti-AKT isoform specific Antibodies were produced by repeated immunizations with a synthetic peptide corresponding to internal residues of human AKT1, AKT2 or AKT3 protein.
• Concentration: 1.0 mg/ml
• Dilution range: ELISA: 1:2,000 - 1:10,000, FC: User Optimized, IHC: 20 µg/mL, WB: 1:1000
• Form/Appearance: Liquid: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
• Purity: This primary antibody kit has been carefully chosen to recognize all 3 isoforms of AKT. Anti-AKT1 antibody is directed against human AKT1. Anti-AKT2 antibody is directed against human AKT2. Anti-AKT3 antibody is directed against human AKT3. The antibodies detect both unphosphorylated and phosphorylated forms of the protein. The antibodies were purified from ascites by Protein A chromatography.
• Conjugation: Unconjugated
• UniProt ID: P31749
• Storage: Each vial contains a relatively low volume of reagent (25 µL). Store vial at -20° C or below prior to opening. To minimize loss of volume dilute 1:10 by adding 225 µL of the buffer stated above directly to the vial. Recap, mix thoroughly and briefly centrifuge to collect the volume at the bottom of the vial. Use this intermediate dilution when calculating final dilutions as recommended below. Store the vial at -20°C or below after dilution. Avoid cycles of freezing and thawing.
• Buffer/Preservatives: 0.01% (w/v) Sodium Azide
• Alternative names: AKT isoform antibody kit, AKT-1, AKT-2, AKT-3, Pro
• Note: For research use only
• Application notes: Anti-AKT Antibodies are suitable for ELISA, immunohistochemistry, and western blotting. Expect a band approximately 56 kDa in size corresponding to AKT proteins by western blotting in the appropriate cell lysate or extract. All 3 primary monoclonal antibodies reacts with human AKT. Specific conditions for reactivity should be optimized by the end user. For immunohistochemistry we recommend the use of fresh frozen tissues. Attempts at staining paraffin-embedded formalin fixed tissues were negative. No pre-treatment of sample is required. This sampler kit consists of sample sizes of anti-AKT1, anti-AKT2 and anti-AKT3 mouse monoclonal antibody and HRP conjugated anti-Mouse IgG secondary antibody (p/n 200-301-I51S; 200-301-BR8S; 200-301-J36S and 610-403-C46S, respectively).
• Expiration Date: Please enquire.

Flow Cytometry of Mouse anti-AKT1 antibody. Cells: LNCap Cells. Stimulation: none. Primary antibody: Allophycocyanin AKT1 antibody at 1.0 µg/mL for 20 min at 4°C.

Immunofluorescence of Mouse monoclonal anti-AKT3 antibody. Cell Type: A431 cells. Fixation: 4% paraformaldehyde 10 min. Permeabilization: 0.5% Triton X 30 min. Primary Ab: (p/n orb344529) at 1:250 for 72 hours 4°C. Secondary Ab: at 1:1000 overnight 4°C.

Plate was coated with monoclonal anti AKT1 antibody (capture antibody) followed by incubation with recombinant AKT1 (p/n orb346473), AKT2 (p/n orb346474), AKT3 (p/n orb346475) proteins. Binding was detected with biotinylated monoclonal anti-AKT pS473. The signal shows specificity of the monoclonal anti-AKT1 antibody to recombinant isoform AKT1 protein and not the isoform 2 and 3.

Western Blot of Mouse Anti-AKT1 antibody. Lane 1: AKT-1 Null. Lane 2: WT. Lane 3: MEF #1. Lane 4 : A549. Lane 5: Calu-1. Lane 6: PC-3 (p/n orb692718). Lane 7: HepG2 (p/n orb348735). Lane 8: Jurkat (p/n orb348674). Lane 9: SKOV3. Lane 10: HEK293T. Lane 11: C2C12 (p/n orb348725). Load: 20 ug per lane. Primary antibody: AKT1 antibody at 1:1000 for overnight at 4°C. Secondary antibody: Peroxidase mouse secondary antibody at 1:40000 for 30 min at RT. Block: orb348637 for 30 min at RT. Predicted/Observed size: 56 kDa for AKT1. Other band(s): none.

Western Blot of Mouse Anti-AKT1 antibody. Lane 1: GST Tagged recombinant AKT1. Lane 2: GST Tagged recombinant AKT2. Lane 3: GST Tagged recombinant AKT3. Load: 25 ng per lane. Primary antibody: AKT1 antibody at 1:1000 for overnight at 4°C. Secondary antibody: Peroxidase mouse secondary antibody at 1:40000 for 30 min at RT. Block: orb348637 for 30 min at RT. Predicted/Observed size: 78 kDa for AKT1. Other band(s): none.