Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283)

Overview

• Product nameAnti-AKT1 (phospho S473) antibody [EP2109Y]See all AKT1 primary antibodies ...
• Description
  Rabbit monoclonal [EP2109Y] to AKT1 (phospho S473)
• Specificityab81283 detects AKT1 phosphorylated at Serine 473.
• Tested applicationsIHC-P, ICC, WB more details
• Species reactivity
  Reacts with: Mouse, Human
  

  Does not react with

  Rat
• Immunogen

  A phospho specific peptide corresponding to residues surrounding Ser473 of human AKT1.

• Positive control3T3 cell lysate treated with PDGF. Cervical carcinoma.
• General notes

  Produced under U.S. Patent No. 5,675,063.

Properties

• FormLiquid
• Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
• Storage bufferPBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%
• PurityTissue culture supernatant
• Clonality Monoclonal
• Clone numberEP2109Y
• IsotypeIgG
• Research Areas
  • Metabolism
  • Pathways and Processes
  • Metabolism processes
  • Apoptosis

  • Cancer
  • Signal transduction
  • Protein phosphorylation
  • Serine/threonine kinases
  • AKT

  • Signal Transduction
  • Protein Phosphorylation
  • Ser / Thr Kinases
  • PKB / AKT

  • Epigenetics and Nuclear Signaling
  • Cell cycle
  • Apoptosis
  • Nuclear

Applications

Target

• FunctionPlays a role as a key modulator of the AKT-mTOR signaling pathway controlling the tempo of the process of newborn neurons integration during adult neurogenesis, including correct neuron positioning, dendritic development and synapse formation (By similarity). 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 by phosphorylating TSC2 at 'Ser-939' and 'Thr-1462', thereby activating mTORC1 signaling and leading to both phosphorylation of 4E-BP1 and in activation of RPS6KB1. Promotes glycogen synthesis by mediating the insulin-induced activation of glycogen synthase. The activated form can suppress FoxO gene transcription and promote cell cycle progression. Essential for the SPATA13-mediated regulation of cell migration and adhesion assembly and disassembly.
• Tissue specificityExpressed in all human cell types so far analyzed. The Tyr-176 phosphorylated form shows a significant increase in expression in breast cancers during the progressive stages i.e. normal to hyperplasia (ADH), ductal carcinoma in situ (DCIS), invasive ductal carcinoma (IDC) and lymph node metastatic (LNMM) stages.
• Involvement in diseaseDefects in AKT1 are a cause of susceptibility to breast cancer (BC) [MIM:114480]. A common malignancy originating from breast epithelial tissue. Breast neoplasms can be distinguished by their histologic pattern. Invasive ductal carcinoma is by far the most common type. Breast cancer is etiologically and genetically heterogeneous. Important genetic factors have been indicated by familial occurrence and bilateral involvement. Mutations at more than one locus can be involved in different families or even in the same case.
  Defects in AKT1 are associated with colorectal cancer (CRC) [MIM:114500].
  Defects in AKT1 are associated with susceptibility to ovarian cancer [MIM:604370]; also called susceptibility to familial breast-ovarian cancer type 1 (BROVCA1).
• Sequence similaritiesBelongs 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.
• DomainBinding of the PH domain to the phosphatidylinositol 3-kinase alpha (PI(3)K) results in its targeting to the plasma membrane. The PH domain mediates interaction with TNK2 and Tyr-176 is also essential for this interaction.
  The AGC-kinase C-terminal mediates interaction with THEM4.
• Post-translational
  modificationsPhosphorylation on Thr-308, Ser-473 and Tyr-474 is required for full activity. Activated TNK2 phosphorylates it on Tyr-176 resulting in its binding to the anionic plasma membrane phospholipid PA. This phosphorylated form localizes to the cell membrane, where it is targeted by PDPK1 and PDPK2 for further phosphorylations on Thr-308 and Ser-473 leading to its activation. Ser-473 phosphorylation by mTORC2 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.
  Ubiquitinated; undergoes both 'Lys-48'- and 'Lys-63'-linked polyubiquitination. TRAF6-induced 'Lys-63'-linked AKT1 ubiquitination is critical for phosphorylation and activation. When ubiquitinated, it translocates to the plasma membrane, where it becomes phosphorylated. When fully phosphorylated and translocated into the nucleus, undergoes 'Lys-48'-polyubiquitination catalyzed by TTC3, leading to its degradation by the proteasome.
• Cellular localizationCytoplasm. Nucleus. Cell membrane. Nucleus after activation by integrin-linked protein kinase 1 (ILK1). Nuclear translocation is enhanced by interaction with TCL1A. Phosphorylation on Tyr-176 by TNK2 results in its localization to the cell membrane where it is targeted for further phosphorylations on Thr-308 and Ser-473 leading to its activation and the activated form translocates to the nucleus.

Target information above from: UniProt accession P31749 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
• Database links
  • Entrez Gene: 207 Human
  • Entrez Gene: 11651 Mouse
  • Omim: 164730 Human
  • SwissProt: P31749 Human
  • SwissProt: P31750 Mouse
  • Unigene: 525622 Human
  • Unigene: 6645 Mouse

• Alternative names
  AKT 1 antibodyAKT antibodyAKT1 antibody

  AKT1_HUMAN antibodyC AKT antibodycAKT antibodyMGC99656 antibodyPKB antibodyPKB-ALPHA antibodyPRKBA antibodyProtein Kinase B Alpha antibodyProtein kinase B antibodyProto-oncogene c-Akt antibodyRAC Alpha antibodyRAC alpha serine/threonine protein kinase antibodyRAC antibodyRAC PK Alpha antibodyRac protein kinase alpha antibodyRAC Serine/Threonine Protein Kinase antibodyRAC-alpha serine/threonine-protein kinase antibodyRAC-PK-alpha antibodyv akt murine thymoma viral oncogene homolog 1 antibodyvAKT Murine Thymoma Viral Oncogene Homolog 1 antibody

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Anti-AKT1 (phospho S473) antibody [EP2109Y] images

• 

  Western blot - AKT1 (phospho S473) antibody [EP2109Y] (ab81283)
  All lanes : Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283) at 1/10000 dilution
  
  Lane 1 : 3T3 cell lysate untreated
  Lane 2 : 3T3 cell lysates treated with PDGF
  
  Lysates/proteins at 10 µg per lane.
  
  Secondary
  HRP conjugated goat anti-rabbit at 1/2000 dilution
  
  Predicted band size : 56 kDa
  Observed band size : 60 kDa (why is the actual band size different from the predicted?)
  
• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - AKT1 (phospho S473) antibody [EP2109Y] (ab81283)
  ab81283, at 1/100 dilution, staining AKT1 in untreated (left panel) and Phosphatase-treated (right panel) cervical carcinoma by Immunohistochemistry using formalin-fixed, paraffin-embedded tissue
• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283)Image from Henken FE et al., Mol Cancer. 2011 Jun 10;10:71. Fig 7.; doi:10.1186/1476-4598-10-71; 10 June 2011, Molecular Cancer 2011, 10:71
  Immunohistochemical analysis of Human HPV16 immortalized keratinocytes transfected with non-targeting siRNA, staining AKT1 (phospho S473) (green) with ab81283.
  
  Antigen retrieval was performed by heat mediation in citrate buffer (pH 6). Samples were blocked with 10% goat serum before incubating with primary antibody (1/100). Fluoroscein-conjugated tyramide was used to detect staining.
• 

  Immunocytochemistry/ Immunofluorescence-Anti-AKT1 (phospho S473) antibody [EP2109Y](ab81283)
  ab81283 staining AKT1 (phospho S473) in MCF7 cells treated with DAPT (ab120633), by ICC/IF. Decrease in expression of AKT1 (phospho S473) correlates with increased concentration of DAPT, as described in literature.
  The cells were incubated at 37°C for 24h in media containing different concentrations of ab120633 (DAPT) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab81283 (1/200 dilution) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.