AKT pS473 antibody (Biotin)

• Catalog Number: orb344541
• Category: Antibodies
• Description: AKT pS473 antibody (Biotin)
• Species/Host: Mouse
• Clonality: Monoclonal
• Clone Number: 17F6.B11
• Tested applications: ELISA, FC, IF, IHC, IP, WB
• Reactivity: Human, Monkey, Mouse, Rat
• Isotype: IgG1
• Immunogen: Anti-AKT pS473 (MOUSE) Monoclonal Antibody was produced by repeated immunizations with a synthetic peptide corresponding to residues surrounding S473 of human AKT1 protein.
• Concentration: 0.5 mg/mL
• Dilution range: ELISA: 1:20,000, FC: User Optimized, IHC: 20 µg/mL, IF: 1:500-1:3,000, IP: User Optimized, WB: 1:500-1:3,000
• Form/Appearance: Lyophilized
• Purity: Anti-AKT pS473 (MOUSE) Monoclonal Antibody Biotin Conjugated was purified from concentrated tissue culture supernate by Protein A chromatography. This antibody is specific for human and mouse AKT protein phosphorylated at S473. A BLAST analysis was used to suggest cross-reactivity with AKT pS473 from human, mouse, rat and chimpanzee sources based on 100% homology with the immunizing sequence. Cross-reactivity with AKT from other sources has not been determined. Cross-reactivity with AKT2 and AKT3 has not been determined.
• Conjugation: Biotin
• UniProt ID: P31749
• NCBI: 62241011
• Storage: Store vial at 4° C prior to restoration. For extended storage aliquot contents and freeze at -20° C or below. 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: mouse anti-AKT pS473 biotin conjugated Antibody, b
• Note: For research use only
• Application notes: Biotin Conjugated Anti-AKT pS473 (MOUSE) Monoclonal Antibody Biotin Conjugated is tested for ELISA, immunohistochemistry, immunoprecipitation and western blotting. Expect a band approximately 56 kDa in size corresponding to phosphorylated AKT protein by western blotting in the appropriate cell lysate or extract. This phospho-specific monoclonal antibody reacts with human and mouse AKT pS473 and shows minimal reactivity by ELISA against the non-phosphorylated form of the immunizing peptide. Specific conditions for reactivity should be optimized by the end user. For immunohistochemistry use formalin-fixed paraffin-embedded sections. No pre-treatment of sample is required.
• Expiration Date: 12 months from date of receipt.

ELISA of Mouse anti-Akt phospho S473 Biotin Conjugated antibody. Antigen: BSA conjugates of Akt phospho S473 and AKT non-phospho S473. Coating amount: 0.1 µg per well. Primary antibody: Akt phospho S473 Biotin Conjugated antibody at 5 µg/mL. Dilution series: 3-fold. Mid-point concentration: 5 ng/mL Akt phospho S473 Biotin Conjugated antibody. Secondary antibody: Peroxidase streptavidin secondary antibody at 1:10000. Substrate: TMB.

Immunohistochemistry of mouse anti AKT phospho S473 biotin conjugated. Tissue: prostate at 40X (left) with negative control (right). Fixation: FFPE buffered formalin 10% conc. Antigen retrieval: Heat, Citrate pH 6.2. Pressure Cooker. Primary antibody: AKT pS473 biotin at 20 µg/mL for 1 h at RT. Secondary antibody: Streptavidin Conj. HRP at 10 ug/ml. Localization: nuclear and occasionally cytoplasmic. Staining: antibody as precipitated red signal with a hematoxylin purple nuclear counterstain.

Immunohistochemistry of mouse Anti-AKT pS473 (MOUSE) Biotin Conjugated. Tissue: prostate at 40X. Fixation: FFPE buffered formalin 10% conc. Antigen retrieval: Heat, Citrate pH 6.2. Pressure Cooker, left. (pH 9 shown on right as negative control). Primary antibody: AKTsS473 biotin 20 µg/mL for 1 h at RT. Secondary antibody: Streptavidin Conj. HRP at 10 ug/ml. Localization: nuclear and occasionally cytoplasmic. Staining: antibody as precipitated red signal with a hematoxylin purple nuclear counterstain.

Western Blot of Mouse anti-Akt phospho S473 Biotin Conjugated antibody. Lane 1: GST tagged AKT1 active recombinant protein. Lane 2: none. Load: 25 ng per lane. Primary antibody: Akt phospho S473 Biotin Conjugated antibody at 1:1000 for overnight at 4°C. Secondary antibody: HRP Streptavidin secondary antibody at 1:40000 for 30 min at RT. Block: orb348637 for 30 min at RT. Predicted/Observed size: 79 kDa, 79 kDa for Akt phospho S473. Other band(s): none.