Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-AKT1 + AKT2 antibody [EPR17062] (ab182729)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR17062] to AKT1 + AKT2
  • Suitable for: IHC-P, WB, ICC/IF, IP, Flow Cyt
  • Reacts with: Mouse, Rat, Human, Recombinant fragment

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 & Species

    Application Species
    Flow Cyt
    Mouse
    Human
    ICC/IF
    Mouse
    Human
    IHC-P
    Mouse
    Rat
    Human
    IP
    Human
    WB
    Mouse
    Rat
    Human
    Recombinant fragment
    See all applications and species data

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • 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

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

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

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR17062

  • Isotype

    IgG

  • Research areas

Images

  • Western blot - Anti-AKT1 + AKT2 antibody [EPR17062] (ab182729)
    Western blot - Anti-AKT1 + AKT2 antibody [EPR17062] (ab182729)
    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®.

  • Western blot - Anti-AKT1 + AKT2 antibody [EPR17062] (ab182729)
    Western blot - Anti-AKT1 + AKT2 antibody [EPR17062] (ab182729)
    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.

  • Western blot - Anti-AKT1 + AKT2 antibody [EPR17062] (ab182729)
    Western blot - Anti-AKT1 + AKT2 antibody [EPR17062] (ab182729)
    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.

  • Western blot - Anti-AKT1 + AKT2 antibody [EPR17062] (ab182729)
    Western blot - Anti-AKT1 + AKT2 antibody [EPR17062] (ab182729)
    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.

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKT1 + AKT2 antibody [EPR17062] (ab182729)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKT1 + AKT2 antibody [EPR17062] (ab182729)

    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKT1 + AKT2 antibody [EPR17062] (ab182729)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKT1 + AKT2 antibody [EPR17062] (ab182729)

    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKT1 + AKT2 antibody [EPR17062] (ab182729)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKT1 + AKT2 antibody [EPR17062] (ab182729)

    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKT1 + AKT2 antibody [EPR17062] (ab182729)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKT1 + AKT2 antibody [EPR17062] (ab182729)

    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.

  • Immunocytochemistry/ Immunofluorescence - Anti-AKT1 + AKT2 antibody [EPR17062] (ab182729)
    Immunocytochemistry/ Immunofluorescence - Anti-AKT1 + AKT2 antibody [EPR17062] (ab182729)

    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.

  • Immunocytochemistry/ Immunofluorescence - Anti-AKT1 + AKT2 antibody [EPR17062] (ab182729)
    Immunocytochemistry/ Immunofluorescence - Anti-AKT1 + AKT2 antibody [EPR17062] (ab182729)

    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 Cytometry - Anti-AKT1 + AKT2 antibody [EPR17062] (ab182729)
    Flow Cytometry - Anti-AKT1 + AKT2 antibody [EPR17062] (ab182729)

    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 Cytometry - Anti-AKT1 + AKT2 antibody [EPR17062] (ab182729)
    Flow Cytometry - Anti-AKT1 + AKT2 antibody [EPR17062] (ab182729)

    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.

  • Immunoprecipitation - Anti-AKT1 + AKT2 antibody [EPR17062] (ab182729)
    Immunoprecipitation - Anti-AKT1 + AKT2 antibody [EPR17062] (ab182729)

    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.

  • Anti-AKT1 + AKT2 antibody [EPR17062] (ab182729)
    Anti-AKT1 + AKT2 antibody [EPR17062] (ab182729)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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