Overview

  • Product name
    Anti-AKT1/2/3 antibody [EPR16798]
    See all AKT1/2/3 primary antibodies
  • Description
    Rabbit monoclonal [EPR16798] to AKT1/2/3
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Xenopus tropicalis
  • Immunogen

    Recombinant fragment within Human AKT1/2/3 aa 250 to the C-terminus. The exact sequence is proprietary. Other SwissProt IDs: P31751, Q9Y243
    Database link: P31749

  • Positive control
    • WB: MCF7, HeLa, Hep G2 and A549 whole cell lysates; Human fetal brain, heart and kidney lysates; Mouse and Rat brain, heart, kidney and spleen lysates; Xenopus muscle lysate; AKT2 and AKT3 recombinant proteins. IHC-P: Human kidney, Mouse and Rat cerebral cortex. ICC/IF: K562 cells. Flow: A549 cells. IP: MCF7 whole cell lysate
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab179463 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/10000. Detects a band of approximately 56 kDa (predicted molecular weight: 56 kDa).
IHC-P 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/100.
Flow Cyt 1/330.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IP 1/100.

Target

  • Function
    IGF-1 leads to the activation of AKT3, which may play a role in regulating cell survival. Capable of phosphorylating several known proteins. Truncated isoform 2/PKB gamma 1 without the second serine phosphorylation site could still be stimulated but to a lesser extent.
  • Tissue specificity
    In adult tissues, it is highly expressed in brain, lung and kidney, but weakly in heart, testis and liver. In fetal tissues, it is highly expressed in heart, liver and brain and not at all in kidney.
  • Sequence similarities
    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.
  • Domain
    Binding of the PH domain to the phosphatidylinositol 3-kinase alpha (PI(3)K) results in its targeting to the plasma membrane.
  • Post-translational
    modifications
    Phosphorylation on Thr-305 and Ser-472 is required for full activity (By similarity). Phosphorylated upon DNA damage, probably by ATM or ATR.
    Ubiquitinated. When fully phosphorylated and translocated into the nucleus, undergoes 'Lys-48'-polyubiquitination catalyzed by TTC3, leading to its degradation by the proteasome.
  • Cellular localization
    Cytoplasm. Membrane. Membrane-associated after cell stimulation leading to its translocation.
  • Information by UniProt
  • Database links
  • Alternative names
    • AKT antibody
    • AKT1 antibody
    • AKT1 kinase antibody
    • AKT1m antibody
    • AKT2 antibody
    • AKT2 kinase antibody
    • Akt3 antibody
    • AKT3_HUMAN antibody
    • CAKT antibody
    • CWS6 antibody
    • DKFZp434N0250 antibody
    • HIHGHH antibody
    • kinase Akt1 antibody
    • MGC99656 antibody
    • MPPH antibody
    • Murine thymoma viral (v-akt) homolog 2 antibody
    • PKB ALPHA antibody
    • PKB antibody
    • PKB beta antibody
    • PKB gamma antibody
    • PKB-GAMMA antibody
    • PKB/Akt antibody
    • PKBALPHA antibody
    • PKBB antibody
    • PKBBETA antibody
    • PKBG antibody
    • PKBGAMMA antibody
    • PRKBA antibody
    • PRKBB antibody
    • PRKBG antibody
    • Protein kinase Akt 2 antibody
    • Protein kinase Akt-3 antibody
    • Protein kinase B alpha antibody
    • Protein kinase B antibody
    • Protein kinase B beta antibody
    • Protein kinase B gamma antibody
    • Proto oncogene c Akt antibody
    • RAC ALPHA antibody
    • RAC alpha serine/threonine protein kinase antibody
    • RAC antibody
    • RAC BETA antibody
    • RAC beta serine/threonine protein kinase antibody
    • RAC PK alpha antibody
    • RAC PK beta antibody
    • rac protein kinase alpha antibody
    • rac protein kinase beta antibody
    • RAC-gamma antibody
    • RAC-gamma serine/threonine-protein kinase antibody
    • RAC-PK-gamma antibody
    • RACALPHA antibody
    • RACalpha serine/threonine kinase antibody
    • RACBETA antibody
    • RACgamma antibody
    • RACgamma serine/threonine protein kinase antibody
    • RACPKgamma antibody
    • serine threonine protein kinase antibody
    • STK-2 antibody
    • STK2 antibody
    • thymoma viral proto oncogene 1 antibody
    • thymoma viral proto oncogene antibody
    • V akt murine thymoma viral oncogene homolog 1 antibody
    • V akt murine thymoma viral oncogene homolog 2 antibody
    • V akt murine thymoma viral oncogene homolog 3 (protein kinase B, gamma) antibody
    • V akt murine thymoma viral oncogene homolog 3 antibody
    • vakt murine thymoma viral oncogene homolog 1 antibody
    • vakt murine thymoma viral oncogene homolog 2 antibody
    • vakt murine thymoma viral oncogene homolog 3 antibody
    see all

Images

  • All lanes : Anti-AKT1/2/3 antibody [EPR16798] (ab179463) at 1/10000 dilution

    Lane 1 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysates
    Lane 2 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysates
    Lane 3 : Hep G2 (Human liver hepatocellular carcinoma) whole cell lysates
    Lane 4 : A549 (Human lung carcinoma) whole cell lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 56 kDa
    Observed band size: 56 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized K562 (Human chronic myelogenous leukemia cells from bone marrow) cells labeling AKT1/2/3 with ab179463 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/400 dilution (green). Cytoplasm and nuclear staining on K562 cell line is observed. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
    The negative controls are as follows;
    1. ab179463 at 1/100 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    2. ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/400 dilution.

     

  • Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling AKT1/2/3 with ab179463 at 1/250 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Cytoplasm and nucleus staining on Human renal cortex is observed. Counter stained with Hematoxylin.

    Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

  • All lanes : Anti-AKT1/2/3 antibody [EPR16798] (ab179463) at 1/1000 dilution

    Lane 1 : Human fetal brain lysate
    Lane 2 : Human fetal heart lysate
    Lane 3 : Human fetal kidney lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

    Predicted band size: 56 kDa
    Observed band size: 56 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-AKT1/2/3 antibody [EPR16798] (ab179463) at 1/1000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Mouse heart lysate
    Lane 3 : Mouse kidney lysate
    Lane 4 : Mouse spleen lysate
    Lane 5 : Rat brain lysate
    Lane 6 : Rat heart lysate
    Lane 7 : Rat kidney lysate
    Lane 8 : Rat spleen lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 56 kDa
    Observed band size: 56 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Anti-AKT1/2/3 antibody [EPR16798] (ab179463) at 1/10000 dilution + Xenopus muscle lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 56 kDa
    Observed band size: 56 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-AKT1/2/3 antibody [EPR16798] (ab179463) at 1/10000 dilution

    Lane 1 : AKT2 recombinant protein (HIS-tag): aa282-481
    Lane 2 : AKT3 recombinant protein (HIS-tag) :aa351-479

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 56 kDa
    Observed band size: 18,26 kDa
    why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunohistochemical analysis of paraffin-embedded Mouse cerebral cortex tissue labeling AKT1/2/3 with ab179463 at 1/250 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Cytoplasm and nucleus staining on Mouse cerebral cortex is observed. Counter stained with Hematoxylin.

    Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

  • Immunohistochemical analysis of paraffin-embedded Rat cerebral cortex tissue labeling AKT1/2/3 with ab179463 at 1/250 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Cytoplasm and nucleus staining on Rat cerebral cortex is observed. Counter stained with Hematoxylin.

    Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

  • Flow cytometric analysis of 2% paraformaldehyde-fixed A549 (Human lung carcinoma) cells labeling AKT1/2/3 with ab179463 at 1/330 dilution (red) compared with a rabbit monoclonal IgG isotype control (black) and a unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.

  • AKT1/2/3 was immunoprecipitated from 1mg of MCF7 (Human breast adenocarcinoma cell line) whole cell extract with ab179463 at 1/100 dilution. Western blot was performed from the immunoprecipitate using ab179463 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution. Lane 1: MCF7 whole cell extract. Lane 2: PBS instead of MCF7 whole cell extract.
    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

References

This product has been referenced in:
  • He F  et al. Low-dose lipopolysaccharide inhibits neuronal apoptosis induced by cerebral ischemia/reperfusion injury via the PI3K/Akt/FoxO1 signaling pathway in rats. Mol Med Rep 19:1443-1452 (2019). Read more (PubMed: 30628689) »
  • Yang X  et al. Downregulation of miR-183 inhibits the growth of PANC-1 pancreatic cancer cells in vitro and in vivo, and increases chemosensitivity to 5-fluorouracil and gemcitabine. Exp Ther Med 17:1697-1705 (2019). Read more (PubMed: 30783438) »
See all 40 Publications for this product

Customer reviews and Q&As

There are currently no Customer reviews or Questions for ab179463.
Please use the links above to contact us or submit feedback about this product.

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up