AKT pS473 antibody

• Catalog Number: orb345378
• Category: Antibodies
• Description: AKT pS473 antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: DOT, ELISA, IF, IHC, Multiplex Assay, WB
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: Rabbit Anti-AKTpS473 Antibody was prepared by repeated immunizations in rabbits with a synthetic peptide corresponding to a C-terminus region near phospho Serine 473 of the human, mouse, rat and chicken AKT proteins conjugated to KLH.
• Concentration: 1.15 mg/mL
• Dilution range: ELISA: 1:15,000 - 1:60,000, IHC: 1:100 - 1:500, IF: User Optimized, WB: 1:200 - 1:1000
• Form/Appearance: Liquid (sterile filtered)
• Purity: This product was prepared from monospecific antiserum by immunoaffinity chromatography using a phospho peptide coupled to agarose beads followed by solid phase adsorption(s) against a non-phospho peptide and non-specific peptide to remove any unwanted reactivities. Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Rabbit Serum. This antibody is specific for phosphorylated human AKTpS473. Minimal reactivity occurs against non-phosphorylated AKT. Reactivity against AKT from other species may occur but has not yet been tested.
• Conjugation: Unconjugated
• UniProt ID: P31749
• NCBI: 62241011
• Storage: Store vial at -20° C prior to opening. Aliquot contents and freeze at -20° C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use.
• Buffer/Preservatives: 0.01% (w/v) Sodium Azide
• Alternative names: rabbit anti-AKT pS473 Antibody, AKT1 phospho S473,
• Note: For research use only
• Application notes: This AKT antibody is phospho specific for pS473 and is tested in ELISA, western blotting, immunohistochemistry (formalin-fixed paraffin-embedded sections), and immunofluorescence. By immunoblot a single band of the expected apparent molecular weight ~56kDa is observed. For immunohistochemistry no pre-treatment of sample is required.
• Expiration Date: 12 months from date of receipt.

Dot Blot of Rabbit Anti-AKT pS473 Antibody. Dilutions in Columns: (1) 100 ng, (2) 33.33 ng, (3) 11.11 ng, (4) 3.7 ng, (5) 1.23 ng. Tested BSA Peptide Reactivity in Rows: (A) AKT1-BSA, (B) AKT1 pT308-BSA, (C) AKT1 S473-BSA, (D) AKT1 pS473-BSA, (E) CDC27 T244-BSA, (F) CDC27 pT244-BSA, (G) BSA control. Primary Antibody: Anti-AKTpS473 at 1 µg/mL overnight at 2-8°C. Secondary Antibody: Goat anti-Rabbit IgG HRP (p/n orb347654) at 1:70000 at RT for 30 mins. Block: BlockOut Buffer (p/n orb348644).

Immunofluorescence Confocal Microscopy of Rabbit anti-AKT pS473 antibody. Tissue: cardiomyocytes infected with adenovirus expressing with wild-type AKT. Fixation: 0.5% PFA. Antigen retrieval: not required. Primary antibody: AKT pS473 antibody at 1:40 for 1 h at RT. Secondary antibody: texas-red conjugated rabbit secondary antibody at 1:10000 for 45 min at RT. Localization: AKT pS473 is nuclear. Staining: AKT pS473 as green fluorescent signal with texas-red conjugated phalloidin (red) to label filamentous actin.

Immunohistochemistry at higher magnification of Rabbit Anti-Akt pS473 antibody. Tissue: human breast carcinoma. Fixation: formalin fixed paraffin embedded. Antigen retrieval: not required. Primary antibody: Akt pS473 antibody at 100 dilution for 1 h at RT. Secondary antibody: Dako's Techmate streptavidin-biotin reagents at 1:10000 for 45 min at RT. Localization: Akt pS473 is nuclear and occasionally cytoplasmic.

Immunohistochemistry of Rabbit anti-AKT pS473 antibody. Tissue: human breast carcinoma. Fixation: formalin fixed paraffin embedded. Antigen retrieval: not required. Primary antibody: AKT pS473 antibody at 1:100 for 1 h at RT. Secondary antibody: Dako's Techmate streptavidin-biotin reagents at 1:10000 for 45 min at RT. Localization: AKT pS473 is nuclear and occasionally cytoplasmic.

Immunohistochemistry of Rabbit Anti-Akt pS473 antibody. Tissue: human breast carcinoma. Fixation: formalin fixed paraffin embedded. Antigen retrieval: not required. Primary antibody: Akt pS473 antibody at 100 dilution for 1 h at RT. Secondary antibody: Dako's Techmate streptavidin-biotin reagents at 1:10000 for 45 min at RT. Localization: Akt pS473 is nuclear and occasionally cytoplasmic.

Western Blot of Rabbit anti-AKT pS473 antibody. Lane 1: AKT1 Recombinant Protein (p/n orb346473). Lane 2: AKT1 Mutant Human Recombinant Protein (p/n orb346474). Lane 3: AKT1 (phosphatase treated) Human Recombinant Protein (p/n orb346472). Load: 50 ng per lane. Primary antibody: AKT pS473 antibody at 1:1000 for overnight at 4°C. Secondary antibody: Peroxidase rabbit secondary antibody (p/n orb347654) at 1:40000 for 30 min at RT. Block: Blocking Buffer for Fluorescent Western Blotting (orb348637) for 30 min at RT. Predicted/Observed size: ~56 kDa for AKTpS473.