AKT1 Western Chemiluminescent Blotting Kit Antibody

• Catalog Number: orb345942
• Category: Antibodies
• Description: AKT1 Western Chemiluminescent Blotting Kit
• Species/Host: Mouse
• Clonality: Monoclonal
• Clone Number: 14E5.A2.B2.H9
• Tested applications: ELISA, IF, WB
• Reactivity: Human
• Isotype: IgG2a
• Dilution range: WB: User Optimized
• Form/Appearance: n/a
• Purity: The kit is designed to detect both unphosphorylated and phosphorylated forms of the protein. Cross reactivity with AKT1 from other species has not been determined, however, the sequence of the immunogen shows 100% identity to human, mouse, and rat, therefore, cross reactivity is expected. Cross-reactivity with AKT2 and AKT3 has not been determined. This kit contains sufficient substrate for up to 5 mini blots at 7.5 x 8 cm2 (1,800 cm2) and is stable for at least 1 year when stored as indicated.
• Conjugation: Unconjugated
• UniProt ID: P31749
• Storage: See kit insert for complete instructions.
• Buffer/Preservatives: Wash buffers MUST NOT contain SODIUM AZIDE or other inhibitors of peroxidase activity!
• Alternative names: AKT 1 western blotting, AKT-1, PKB antibody, PKB g
• Note: For research use only
• Application notes: Use Biorbyt’ Anti-AKT1 Chemiluminescent Kit for Western Blotting for detection of human AKT1 native and recombinant proteins by western blot. Expect a band approximately 56 kDa in size corresponding to AKT1 protein. This kit is useful for both western blotting and dot blotting methods. Please read the entire product insert prior to use.
• Expiration Date: Please enquire.

PAGE-MAP (microsphere affinity proteomics) of Mouse Anti-AKT1 Antibody. (Catalog Number: orb344523). Antibody array western blot binding of gelfree size separated fractions of multiple lysates (solid lines) and shotgun mass spectroscopy identification (dashed lines) of the target band run in parallel correlate confirming the specificity of this antibody against AKT1.

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.

Western Blot of Mouse Anti-AKT1 Antibody. Lane 1: Opal Prestained Molecular Weight Protein. Lane 2: AKT1 protein (p/n orb346473). Lane 3: AKT2 protein (p/n orb346474). Lane 4: AKT3 protein (p/n orb346475). Load: 50 ng. Blocking: BlockOut Buffer (p/n orb348644) for 30 min at RT. Primary Antibody: Anti-AKT1 at 1 ug/ml o/n at 4°C. Secondary Antibody: Rabbit Anti-Mouse IgG HRP (p/n orb347506) at 1:40000 in orb348644 for 30 min at RT.