Anti-AK-1，腺苷酸激酶-1

• Product nameAnti-AKT1 （phospho S473） antibodySee all AKT1 primary antibodies ...
• Description
  Rabbit polyclonal to AKT1 （phospho S473）
• SpecificityThis antibody does not cross react with non-phosphorylated AKT1 or with other unrelated phosphorylated serines. It may also cross react with phosphorylated AKT2 （S473） ａnd AKT3 （S473） based on sequence homology.
• Tested applicationsIHC-P, ICC/IF, ELISA, Dot Blot, IHC-Fr, WB, IP more details
• Species reactivity
  Reacts with: Mouse, Rat, Human
  Predicted to work with: Chicken, Dog
• Immunogen

  Synthetic phosphopeptide （human） containing S473 from AKT1. （QFS^pYS）

• Positive controlWhole cell lysate from PDGF stimulated 3T3 cells. This antibody gave a positive signal in the following Methanol fixed cell lines: MCF-7.

Properties

• FormLiquid
• Storage instructionsShipped at 4°C. Upon delivery aliquot ａnd store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
• Storage bufferPreservative: None
  Constituents: Tris buffered saline, pH 7.2, containing antibody stabilizer.
• Concentration200 µl at 0.5 mg/ml 
  
• PurityImmunogen affinity purified
• ClonalityPolyclonal
• IsotypeIgG
• Research Areas
  • Signal Transduction
   
  • Protein Phosphorylation
   
  • Ser / Thr Kinases
  • PKB / AKT

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

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

  • Metabolism
   
  • Pathways ａnd Processes
   
  • Metabolism processes
  • Apoptosis

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.
• Information by UniProt
• Database links
  • Entrez Gene: 395928 Chicken
  • Entrez Gene: 490878 Dog
  • Entrez Gene: 207 Human
  • Entrez Gene: 11651 Mouse
  • Entrez Gene: 24185 Rat
  • Omim: 164730 Human
  • SwissProt: P31749 Human
  • SwissProt: P31750 Mouse

  see all
• Alternative names
  • AKT 1 antibody
  • AKT antibody
  • AKT1 antibody

  see all

Anti-AKT1 （phospho S473） antibody images

• 

  Dot Blot - Anti-AKT1 （phospho S473） antibody （ab66138）
  1µg peptide blotted onto a nitrocellulose membrane, followed by ab66138 at 1/2000 dilution. A: AKT1 （pS473） peptide B: AKT1 non-phosphopeptide C: non-related phosphopeptide
• 

  Western blot - AKT1 （phospho S473） antibody （ab66138）
  All lanes : Anti-AKT1 （phospho S473） antibody （ab66138） at 1/1000 dilution
  
  Lane 1 : Whole cell lysate derived from PDGF stimulated 3T3
  Lane 2 : Whole cell lysate derived from PDGF stimulated 3T3 with immunising peptide
  
  Lysates/proteins at 10 µg per lane.
  
  
  Predicted band size : 56 kDa
  Observed band size : 60 kDa （why is the actual band size different from the predicted?）
  
• 

  Immunocytochemistry/ Immunofluorescence - AKT1 （phospho S473） antibody （ab66138）This image is courtesy of an anonymous Abreview.
  ab66138 staining AKT1 in Mouse embryonic stem cells by Immunocytochemistry/Immunofluorescence. The cells were 4% PFA fixed ａnd permeabilized in 0.1% Triton prior to blocking in 10% serum for 30 minutes at room temperature. The primary antibody was diluted 1/1000 ａnd incubated with the sample for 12 hours at 4°C. The secondary antibody was an Alexa Fluor® 594-conjugated Goat anti-Rabbit polyclonal, diluted 1/1500.
• 

  Immunohistochemistry （Frozen sections） - AKT1 （phospho S473） antibody （ab66138）This image is courtesy of an anonymous Abreview
  ab66138 staining phospho AKT1 in Mouse embryonic tissue sections by Immunohistochemistry （IHC-Fr - frozen sections）. Tissue was fixed with paraformaldehyde, permeabilized with 0.1% Triton ａnd blocked with 10% serum for 1 hour at 22°C. Samples were incubated with primary antibody （1/500 in blocking buffer （1/10）） for 16 hours at 4°C. An Alexa Fluor^®488-conjugated Goat anti-rabbit polyclonal （1/1000） was used as the secondary antibody.

  See Abreview

• 

  Western blot - Anti-AKT1 （phospho S473） antibody （ab66138）
  
  developed using the ECL technique
  
  Performed under reducing conditions.
  
  Predicted band size : 56 kDa
  

  PC12 cells were incubated at 37°C for 30 minutes with vehicle control （0 µM） ａnd different concentrations of cabergoline （ab120564）. Increased expression of AKT1 （phospho S473） （ab66138） in PC12 cells correlates with an increase in cabergoline concentration, as described in literature.

   

  Whole cell lysates were prepared with RIPA buffer （containing protease inhibitors ａnd sodium orthovanadate）, 10µg of each were loaded on the gel ａnd the WB was run under reducing conditions. After transfer the membrane was blocked for an hour using 5% BSA before being incubated with ab66138 at 1/1000 dilution and ab8227 at 1 µg/ml overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP （ab97051） at 1/10000 dilution ａnd visualised using ECL development solution.

   

• 

  Immunocytochemistry/ Immunofluorescence - Anti-AKT1 （phospho S473） antibody （ab66138）

  ICC/IF image of ab66138 stained MCF-7 cells. The cells were methanol fixed （5 min） ａnd then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells ａnd block non-specific protein-protein interactions. The cells were then incubated with the antibody ab66138 at 1µg/ml overnight at +4°C. The secondary antibody （green） was DyLight® 488 goat anti- rabbit （ab96899） IgG （H+L） used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes （red） at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei （blue） at a concentration of 1.43µM.

References for Anti-AKT1 （phospho S473） antibody （ab66138）