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Key Specifications Table
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| H, M, R, Vrt | ELISA, WB, FC, IF, IHC | M | Purified | Monoclonal Antibody |
| Description | |
|---|---|
| Catalogue Number | 05-1003 |
| Description | Anti-phospho-Akt (Ser473) Antibody, clone 6F5 |
| Alternate Names |
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| Background Information | 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. |
| Product Information | |
|---|---|
| Format | Purified |
| Presentation | Purified mouse monolconal IgG1κ in 0.1M Tris-Glycine (pH 7.4), 150mM NaCl with 0.05%NaN3 |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| 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. |
| Key Applications |
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| Application Notes | 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-AlexaFluor®488 (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. |
| Biological Information | |
|---|---|
| Immunogen | Peptide encompassing and including phosphorylated Ser473 of human Akt1. |
| Epitope | Phosphorylated Ser473 |
| Clone | 6F5 |
| Concentration | Please refer to the Certificate of Analysis for the lot-specific concentration. |
| Host | Mouse |
| Specificity | Recognizes Akt only when phosphorylated on Ser473. This antibody does not cross-react with Akt1/PKBα when it is not phosphorylated on Ser473. |
| Isotype | IgG1κ |
| Species Reactivity |
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| Species Reactivity Note | Human, mouse, and rat tested. Expected to react with most species as sequence is 100% homologous with most common vertebrate. |
| Antibody Type | Monoclonal Antibody |
| Entrez Gene Number |
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| Entrez Gene Summary | The serine-threonine protein kinase encoded by the AKT1 gene is catalytically inactive in serum-starved primary and immortalized fibroblasts. AKT1 and the related AKT2 are activated by platelet-derived growth factor. The activation is rapid and specific, and it is abrogated by mutations in the pleckstrin homology domain of AKT1. It was shown that the activation occurs through phosphatidylinositol 3-kinase. In the developing nervous system AKT is a critical mediator of growth factor-induced neuronal survival. Survival factors can suppress apoptosis in a transcription-independent manner by activating the serine/threonine kinase AKT1, which then phosphorylates and inactivates components of the apoptotic machinery. Multiple alternatively spliced transcript variants have been found for this gene. [provided by RefSeq]. |
| Gene Symbol |
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| Modifications |
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| Purification Method | Protein G Purified |
| UniProt Number |
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| UniProt Summary | FUNCTION: General protein kinase capable of phosphorylating several known proteins. Phosphorylates TBC1D4. Signals downstream of phosphatidylinositol 3-kinase (PI(3)K) to mediate the effects of various growth factors such as platelet-derived growth factor (PDGF), epidermal growth factor (EGF), insulin and insulin-like growth factor I (IGF-I). Plays a role in glucose transport by mediating insulin-induced translocation of the GLUT4 glucose transporter to the cell surface. Mediates the antiapoptotic effects of IGF-I. Mediates insulin-stimulated protein synthesis, partly by playing a role in both insulin-induced phosphorylation of 4E-BP1 and in insulin-induced activation of p70 S6 kinase. Promotes glycogen synthesis by mediating the insulin-induced activation of glycogen synthase. Ref.4 Ref.8 Ref.12 Ref.13 Ref.16 Ref.20 Catalytic activity ATP + a protein = ADP + a phosphoprotein. Ref.4 Ref.20 Ref.1 ENZYME REGULATION:Three specific sites, one in the kinase domain (Thr-308) and the two other ones in the C-terminal regulatory region (Ser-473 and Tyr-474), need to be phosphorylated for its full activation. Ref.8 Ref.6 SUBUNIT STRUCTURE: Interacts with AGAP2 isoform 2 (PIKE-A) in the presence of guanine nucleotides. The C-terminus interacts with CCDC88A/GRDN and THEM4. Interacts with AKTIP. Interacts (via PH domain) with MTCP1, TCL1A AND TCL1B. Interacts with CDKN1B; the interaction phosphorylates CDKN1B promoting 14-3-3 binding and cell-cycle progression. Ref.13 Ref.16 Ref.20 Ref.9 Ref.10 Ref.11 Ref.15 Ref.17 Ref.18 Ref.19 SUBCELLULAR LOCATION: Cytoplasm. Nucleus. Cell membrane. Note= Nucleus after activation by integrin-linked protein kinase 1 (ILK1). Nuclear translocation is enhanced by interaction with TCL1A. Ref.11 TISSUE SPECIFICTY: In all human cell types so far analyzed. Ref.4 DOMAIN: Binding of the PH domain to the phosphatidylinositol 3-kinase alpha (PI(3)K) results in its targeting to the plasma membrane. The AGC-kinase C-terminal mediates interaction with THEM4. PTM: Phosphorylation on Thr-308, Ser-473 and Tyr-474 is required for full activity. Ser-473 phosphorylation by the Rictor-mTor complex favors Thr-308 phosphorylation by PDPK1. Ser-473 phosphorylation is enhanced by interaction with AGAP2 isoform 2 (PIKE-A). Ser-473 phosphorylation is enhanced in focal cortical dysplasias with Taylor-type balloon cells. SIMILARITY: Belongs to the protein kinase superfamily. AGC Ser/Thr protein kinase family. RAC subfamily. Contains 1 AGC-kinase C-terminal domain. Contains 1 PH domain. Contains 1 protein kinase domain. |
| Molecular Weight | 60 kDa |
| Product Usage Statements | |
|---|---|
| Quality Assurance | 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. |
| Usage Statement |
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| Storage and Shipping Information | |
|---|---|
| Storage Conditions | Stable for 1 year at 2-8ºC from date of receipt. |
| Packaging Information | |
|---|---|
| Material Size | 100 µg |