Description
General description
60 kDa
Akt/PKB is a Ser/Thr kinase and a major known effecter of the PI3 Kinase pathway. It is involved in multiple signaling pathways that relate to many biological processes including glucose metabolism, cell survival/apoptosis, cell cycle control, angiogenesis, differentiation, and cell growth and proliferation. In mammals three isoforms of Akt (Akt1/PKB, Akt2/PKB, and Akt3/PKB)exists. They exhibit a high degree of homology, but differ slightly in the localization of their regulatory phosphorylation sites. Akt is the predominant isoform that is in most tissues and is thought to have a dominant role in growth, survival, embryonic development, and adipocyte differentiation. Akt2 is correlated with the regulation of glucose homeostasis and is the predominant isoform expressed in insulin-responsive tissues. Akt3 is abundant in brain tissue. Each Akt isoform is composed of three functionally distinct regions: an N-terminal Pleckstrin Homology (PH) domain that provides a lipid-binding module to direct Akt to PIP3 at the cell membrane that is necessary for its activation, a central catalytic domain containing Thr308, and a C-terminal hydrophobic motif containing Ser473. The activation of Akt is dependent on a dual regulatory mechanism that requires both its translocation to the plasma membrane and dual phosphorylation on Thr308 and Ser473 by PDK1 and the TORC2 complex, respectively.
Immunogen
Peptide encompassing and including phosphorylated Ser473 of human Akt1.
Application
Anti-phospho-Akt (Ser473) Antibody, clone 6F5 detects level of phospho-Akt (Ser473) & has been published & validated for use in ELISA, WB, FC, IF, IH.
Immunofluroescence Analysis:
EGF-treated A431 cells were fixed, permeabilized, and stained with Anti-phospho-Akt (Ser473), clone 6F5 (2 μg/mL) (red). Cy3-conjugated anti-mouse was used for detection and Phalloidin-AlexaFluor488 (green) was used to stain actin.
Immunohistochemistry:
Anti-phospho-Akt (Ser473), clone 6F5 was diluted to 1:200, IHC-Select Detection with HRP-DAB.
Flow Cytometry:
Anti-phospho-Akt (Ser473) was used at a 1:800 dilution for this assay.
Biochem/physiol Actions
Expected to react with most species as sequence is 100% homologous with most common vertebrate.
Recognizes Akt only when phosphorylated on Ser473. This antibody does not cross-react with Akt1/PKBα when it is not phosphorylated on Ser473.
Physical form
Format: Purified
Purified mouse monolconal IgG1κ in 0.1M Tris-Glycine (pH 7.4), 150mM NaCl with 0.05%NaN3
Analysis Note
Western Blot Analysis:
0.5-1 μg/mL of this lot detected phosphorylated Akt in lysates from mouse NIH-3T3 fibroblasts treated with 100 ng/mL PDGF for 20 minutes.
Other Notes
Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.
