Overview

  • Product name

    Anti-AKT1 + AKT2 antibody [EPR18405]
    See all AKT1 + AKT2 primary antibodies
  • Description

    Rabbit monoclonal [EPR18405] to AKT1 + AKT2
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, WB, Flow Cyt, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    corresponding to Human AKT1 + AKT2 aa 400 to the C-terminus. Also SwissProt: P31751
    Database link: P31749

  • Positive control

    • WB: AKT1 and AKT2 recombinant protein; HeLa, HepG2, A549, MCF-7, PC12 and NIH/3T3 whole cell lysate; Human fetal kidney lysate; Mouse brain lysate, Rat brain and heart lysate. IHC-P: Human kidney, Human gastric adenocarcinoma, Mouse liver and Rat kidney tissue. ICC/IF: HeLa and NIH/3T3 cells. Flow Cytometry: HeLa cells.
  • 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

    EPR18405
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab188099 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 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/2000. Detects a band of approximately 56 kDa (predicted molecular weight: 56 kDa).
Flow Cyt 1/200.
ICC/IF 1/500.

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 [EPR18405] (ab188099) at 1/20000 dilution

    Lane 1 : AKT1 recombinant protein fragment (His-Tag®): aa250-481
    Lane 2 : AKT2 recombinant protein fragment (His-Tag®): aa282-481
    Lane 3 : AKT3 recombinant protein fragment (His-Tag®): aa351-479

    Lysates/proteins at 0.01 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution

    Predicted band size: 56 kDa


    Exposure time: 10 seconds


    Blocking/dilution buffer: 5% NFDM/TBST.

     

    All three Human AKT recombinant protein fragments containing an N-terminal His-Tag® were made in house.

  • All lanes : Anti-AKT1 + AKT2 antibody [EPR18405] (ab188099) at 1/2000 dilution

    Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
    Lane 2 : MCF-7 (Human breast adenocarcinoma) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution

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


    Exposure time: 5 seconds


    Blocking/dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-AKT1 + AKT2 antibody [EPR18405] (ab188099) at 1/2000 dilution

    Lane 1 : HepG2 (Human liver hepatocellular carcinoma) whole cell lysate
    Lane 2 : A549 (Human lung carcinoma) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution

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


    Exposure time: 15 seconds


    Blocking/dilution buffer: 5% NFDM/TBST.

  • Anti-AKT1 + AKT2 antibody [EPR18405] (ab188099) at 1/2000 dilution + Human fetal kidney lysate at 10 µg

    Secondary
    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


    Exposure time: 3 minutes


    Blocking/dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-AKT1 + AKT2 antibody [EPR18405] (ab188099) at 1/2000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Rat brain lysate
    Lane 3 : Rat heart lysate
    Lane 4 : PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate
    Lane 5 : NIH/3T3 (mouse embryo fibroblast) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution

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


    Exposure time: 15 seconds


    Blocking/dilution buffer: 5% NFDM/TBST.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling AKT1 + AKT2 with ab188099 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green).

    Confocal image showing cytoplasmic and weakly nuclear staining on HeLa cells.

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
    The negative controls are as follows:-
    -ve control 1 - ab188099 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    -ve control 2. - ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 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 ab188099 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green).

    Confocal image showing cytoplasmic and nuclear staining on NIH/3T3 cells.

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
    The negative controls are as follows:-
    -ve control 1 - ab188099 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    -ve control 2. - ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

  • Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling AKT1 + AKT2 with ab188099 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
    Nucleus and cytoplasm staining on normal Human kidney is observed.
    Counter stained with Hematoxylin.
    Negative control: Used PBS instead of primary ab, secondary ab is Goat Anti-Rabbit IgG H&L (HRP) (ab97051).

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

  • Immunohistochemical analysis of paraffin-embedded Human gastric adenocarcinoma tissue labeling AKT1 + AKT2 with ab188099 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Nucleus and cytoplasm staining on tumor cells of gastric adenocarcinoma is observed.

    Counter stained with Hematoxylin.
    Negative control: Used PBS instead of primary ab, secondary ab is Goat Anti-Rabbit IgG H&L (HRP) (ab97051).

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

  • Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling AKT1 + AKT2 with ab188099 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Nucleus and cytoplasm staining on hepatocytes of mouse liver is observed.

    Counter stained with Hematoxylin.
    Negative control: Used PBS instead of primary ab, secondary ab is Goat Anti-Rabbit IgG H&L (HRP) (ab97051).

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

  • Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling AKT1 + AKT2 with ab188099 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Nucleus and cytoplasm staining on rat kidney is observed.

    Counter stained with Hematoxylin.
    Negative control: Used PBS instead of primary ab, secondary ab is Goat Anti-Rabbit IgG H&L (HRP) (ab97051).

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

  • Flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling AKT1/2 with ab188099 at 1/200 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730, black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/500 dilution was used as the secondary antibody.

References

This product has been referenced in:

  • Xie X  et al. Fe3O4-solamargine induces apoptosis and inhibits metastasis of pancreatic cancer cells. Int J Oncol 54:905-915 (2019). Read more (PubMed: 30483763) »
See 1 Publication for this product

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