Akt (phospho-T308) antibody

• Catalog Number: orb344406
• Category: Antibodies
• Description: Akt (phospho-T308) antibody
• Species/Host: Mouse
• Clonality: Monoclonal
• Clone Number: 18F3.H11
• Tested applications: ELISA, FC, IF, IHC, IP, WB
• Reactivity: Human, Mouse
• 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, IF: 1:25, IP: User Optimized, WB: 1:500 - 1:3,000
• Form/Appearance: Liquid (sterile filtered)
• Purity: Anti-AKT Phospho pT308 antibody 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: Unconjugated
• UniProt ID: P31749
• NCBI: 62241011
• Storage: Store vial at -20° C or below prior to opening. This vial contains a relatively low volume of reagent (25 µL). 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: mouse anti-AKT pT308 Antibody, RAC-PK-alpha, Prote
• Note: For research use only
• Application notes: Anti-AKT pT308 antibody 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.

Immunohistochemistry of Mouse anti-AKT pT308 antibody. Tissue: human brain cerebellum tissue (40X). Fixation: formalin fixed paraffin embedded. Antigen retrieval: not required. Primary antibody: AKT pT308antibody at 20 µg/mL for 1 h at RT. Secondary antibody: Peroxidase rabbit secondary antibody at 1:10000 for 45 min at RT. Localization: staining of Purkinje neurons and cell processes in the cerebellum, cytosolic as well as occasionally nuclear. Staining: AKT pT308 as precipitated red signal with hematoxylin purple nuclear counterstain.

Western Blot of Mouse anti-AKT pT308 antibody. Lane 1: non-phosphorylated AKT in untreated NIH/3T3 cells (p/n orb348714). Lane 2: phosphorylated AKT on PDGF stimulated NIH/3T3 cell lysates (p/n orb348723). Load: 15 µg per lane. Primary antibody: AKT pT308 antibody at a 1:4000 dilution in TBS with 3% BSA, for 3 h at 4°C. Secondary antibody: Peroxidase conjugated Gt-a-Mouse IgG (Fc) (p/n orb347428) was used at a 1:40000 dilution for 1 h at 4°C. Block: 3% BSA in TBS for 30 min at RT. Predicted/Observed size: (indicated by arrowhead at ~56 kDa). Other band(s): unspecific.