AKT pT308 antibody (Biotin)

• Catalog Number: orb344542
• Category: Antibodies
• Description: AKT pT308 antibody (Biotin)
• Species/Host: Mouse
• Clonality: Monoclonal
• Clone Number: 18F3.H11
• Tested applications: DOT, ELISA, FC, IHC, IP, WB
• Reactivity: Human, Monkey, Mouse, Rat
• Isotype: IgG1
• Immunogen: Anti-AKT pT308 monoclonal antibody was produced by repeated immunizations with a synthetic peptide corresponding to residues surrounding T308 of human AKT1 protein.
• Concentration: 1.0 mg/mL
• Dilution range: ELISA: 1:20,000, FC: User Optimized, IHC: 20 µg/ml, IP: User Optimized, WB: 1:500 - 1:3,000
• Form/Appearance: Lyophilized
• Purity: Anti-AKT pT308 was purified from concentrated tissue culture supernate by Protein A chromatography. This antibody is specific for human and mouse AKT protein phosphorylated at T308. A BLAST analysis was used to suggest cross-reactivity with AKT pT308 from most vertebrate species 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 will likely occur. Cancer, Cell Signaling, Neuroscience, Signal Transduction research.
• 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 pT308 Biotin conjugated Antibody, B
• Note: For research use only
• Application notes: Biotin Conjugated Anti-AKT pT308 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 pT308 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. Use formalin-fixed paraffin-embedded sections for immunohistochemistry. No pre-treatment of sample is required.
• Expiration Date: 12 months from date of receipt.

ELISA of Mouse anti-Akt phospho T308 Biotin Conjugated antibody. Antigen: Unconjugated Akt phospho T308 and AKT non-phospho T308. Coating amount: 0.1 µg per well. Primary antibody: Akt phospho T308 Biotin Conjugated antibody at 5 µg/mL. Dilution series: 3-fold. Mid-point concentration: 5 ng/mL Akt phospho T308 Biotin Conjugated antibody. Secondary antibody: Peroxidase streptavidin secondary antibody at 1:10000. Substrate: TMB.

Immunohistochemistry of Mouse Anti-AKT phospho T308 Biotin Conjugated. Tissue: prostate at 20X (left) and 40X (right). Fixation: FFPE buffered formalin 10% conc. Antigen retrieval: Heat, Citrate pH 6.2. Pressure Cooker, Heat, EDTA pH 9.5 Pressure Cooker. Primary antibody: AKT pT308 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.

Western Blot of Mouse anti-Akt phospho T308 Biotin Conjugated antibody. Lane 1: GST tagged AKT1 active recombinant protein. Lane 2: GST tagged AKT1 un-active recombinant protein. Load: 25 ng per lane. Primary antibody: Akt phospho T308 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 T308. Other band(s): none.