Recombinant
RabMAb

Recombinant Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] - Low endotoxin, Azide free (ab214167)

Overview

  • Product name

    Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] - Low endotoxin, Azide free
    See all AKT1 + AKT2 + AKT3 primary antibodies
  • Description

    Rabbit monoclonal [EPR16798] to AKT1 + AKT2 + AKT3 - Low endotoxin, Azide free
  • 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 + AKT2 + AKT3 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

    ab214167 is a carrier-free antibody designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our Low endotoxin, azide-free formats have low endotoxin level (≤ 1 EU/ml, determined by the LAL assay) and are free from azide, to achieve consistent experimental results in functional assays.

    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 ab214167 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 Use at an assay dependent concentration. Detects a band of approximately 56 kDa (predicted molecular weight: 56 kDa).
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IP Use at an assay dependent concentration.

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
    • pan-akt 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-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
    • 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-ALPHA antibody
    • RAC-alpha serine/threonine-protein kinase antibody
    • RAC-beta serine/threonine-protein kinase antibody
    • RAC-gamma antibody
    • RAC-gamma serine/threonine-protein kinase antibody
    • RAC-PK-alpha antibody
    • RAC-PK-beta 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
    • 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
    • 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 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

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized K562 (Human chronic myelogenous leukemia cells from bone marrow) cells labeling AKT1 + AKT2 + AKT3 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.

     

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab179463).

  • Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling AKT1 + AKT2 + AKT3 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab179463).

    Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded Mouse cerebral cortex tissue labeling AKT1 + AKT2 + AKT3 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab179463).

    Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded Rat cerebral cortex tissue labeling AKT1 + AKT2 + AKT3 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab179463).

    Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Flow cytometric analysis of 2% paraformaldehyde-fixed A549 (Human lung carcinoma) cells labeling AKT1 + AKT2 + AKT3 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab179463).

  • AKT1 + AKT2 + AKT3 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab179463).

References

ab214167 has not yet been referenced specifically in any publications.

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