Overview

  • Product name

    Anti-AKT1 + AKT2 antibody [EPR17062]
    See all AKT1 + AKT2 primary antibodies
  • Description

    Rabbit monoclonal [EPR17062] to AKT1 + AKT2
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, WB, ICC/IF, IP, Flow Cytmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human AKT1 + AKT2 aa 250 to the C-terminus. The exact sequence is proprietary.
    Database link: P31751

  • Positive control

    • WB: Human AKT1 fragment recombinant protein; Human AKT2 fragment recombinant protein; Human fetal heart and fetal kidney lysates; HeLa, MCF7, C6, RAW 264.7 and NIH/3T3 whole cell lysates; Mouse and rat brain and heart lysates. IHC-P: Human tonsil, Human bladder cancer, mouse stomach and rat cerebrum tissues. ICC/IF: HeLa and NIH/3T3 cells. Flow Cyt: HeLa and NIH/3T3 cells. IP: HeLa 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

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR17062
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab182729 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
IHC-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/5000. Detects a band of approximately 56 kDa (predicted molecular weight: 56 kDa).
ICC/IF 1/500.
IP 1/40.
Flow Cyt 1/600.

Target

  • Relevance

    The serine/threonine kinase AKT (protein kinase B or PKB) has a central role in the regulation of several signaling pathways controlling cell proliferation, apoptosis, angiogenesis, and diabetes. In humans, there are three genes in the "AKT family": AKT1, AKT2, and AKT3. AKT1 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. 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. AKT2 is a putative oncogene and is a general protein kinase capable of phophorylating several known proteins. AKT2 is amplified and overexpressed in some human carcinomas. AKT2 acts primarily as a regulator of glucose metabolism.
  • Cellular localization

    ATK1: Cytoplasm. Nucleus. Cell membrane. Note: Nucleus after activation by integrin-linked protein kinase 1 (ILK1). Nuclear translocation is enhanced by interaction with TCL1A.
  • Database links

  • Alternative names

    • AKT 1 antibody
    • AKT 2 antibody
    • AKT antibody
    • AKT1 antibody
    • AKT1 kinase antibody
    • AKT1m antibody
    • AKT2 antibody
    • AKT2 kinase antibody
    • C AKT antibody
    • CWS6 antibody
    • HIHGHH antibody
    • MGC99656 antibody
    • Murine thymoma viral (v akt) homolog 2 antibody
    • Oncogene AKT1 antibody
    • PKB alpha antibody
    • PKB antibody
    • PKB beta antibody
    • PKBB antibody
    • PKBBETA antibody
    • PRKBA antibody
    • PRKBB antibody
    • Protein kinase Akt 2 antibody
    • Protein Kinase B Alpha antibody
    • Protein kinase B antibody
    • Protein kinase B beta antibody
    • Proto oncogene c Akt antibody
    • Proto-oncogene c-Akt antibody
    • RAC antibody
    • RAC alpha antibody
    • RAC alpha serine/threonine protein kinase 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-ALPHA antibody
    • RACbeta 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 1 antibody
    • V-AKT murine thymoma viral oncogene homolog 2 antibody
    see all

Images

  • All lanes : Anti-AKT1 + AKT2 antibody [EPR17062] (ab182729) at 1/1000 dilution

    Lane 1 : Human AKT1 fragment recombinant protein
    Lane 2 : Human AKT2 fragment recombinant protein
    Lane 3 : Human AKT3 fragment recombinant protein

    Lysates/proteins at 0.02 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 56 kDa


    Exposure time: 1 second


    Blocking/Dilution buffer: 5% NFDM/TBST.

    Human AKT1 fragment recombinant protein contains aa281-480 with a His-Tag®. Human AKT2 fragment recombinant protein contains aa282-481 with a His-Tag®. Human AKT3 fragment recombinant protein contains aa351-479 with a His-Tag®.

  • All lanes : Anti-AKT1 + AKT2 antibody [EPR17062] (ab182729) at 1/5000 dilution

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

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

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


    Exposure time: 30 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-AKT1 + AKT2 antibody [EPR17062] (ab182729) at 1/5000 dilution

    Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 2 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

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



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1: 8 seconds; Lane 2: 2 seconds.

  • All lanes : Anti-AKT1 + AKT2 antibody [EPR17062] (ab182729) at 1/5000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Mouse heart lysate
    Lane 3 : Rat brain lysate
    Lane 4 : Rat heart lysate
    Lane 5 : C6 (Rat glial tumor cell line) whole cell lysate
    Lane 6 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
    Lane 7 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

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



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1 2,3 and 4: 5 seconds; Lane 5,6 and 7: 3 seconds.

     

  • Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling AKT1 + AKT2 with ab182729 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm and nuclear staining on Human tonsil is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded Human bladder cancer tissue labeling AKT1 + AKT2 with ab182729 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm and nuclear staining on Human bladder cancer is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded Mouse stomach tissue labeling AKT1 + AKT2 with ab182729 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm and nuclear staining on Mouse stomach is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling AKT1 + AKT2 with ab182729 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm and nuclear staining on Rat cerebrum is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling AKT1 + AKT2 with ab182729 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear and cytoplasmic staining on HeLa cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab150077 at 1/1000 dilution.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling AKT1 + AKT2 with ab182729 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear and cytoplasmic staining on NIH/3T3 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab150077 at 1/1000 dilution.

  • Flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling AKT1 + AKT2 with ab182729 at 1/600 dilution (red) compared with a Rabbit IgG,monoclonal [EPR25A]-Isotype control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti Rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.

  • Flow cytometric analysis of 4% paraformaldehyde-fixed NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling AKT1 + AKT2 with ab182729 at 1/600 dilution (red) compared with a Rabbit IgG,monoclonal [EPR25A]-Isotype control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti Rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.

  • AKT1/2 was immunoprecipitated from 0.35 mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab182729 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab182729 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: HeLa whole cell lysate, 10µg (Input).

    Lane 2: ab182729 IP in HeLa whole cell lysate.

    Lane 3: Rabbit IgG,monoclonal [EPR25A]-Isotype Control (ab172730) instead of ab182729 in HeLa whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 30 seconds.

References

This product has been referenced in:

  • Li J & Zheng J Theaflavins prevent cartilage degeneration via AKT/FOXO3 signaling in vitro. Mol Med Rep 19:821-830 (2019). Read more (PubMed: 30569095) »
  • Li F  et al. CEP55 promotes cell proliferation and inhibits apoptosis via the PI3K/Akt/p21 signaling pathway in human glioma U251 cells. Oncol Lett 15:4789-4796 (2018). Read more (PubMed: 29552118) »
See all 8 Publications for this product

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