重组
RabMAb

Anti-AKT3 + AKT2 + AKT1抗体[Y89] (ab32505)

概述

  • 产品名称
    Anti-AKT3 + AKT2 + AKT1抗体[Y89]
  • 描述
    兔单克隆抗体[Y89] to AKT3 + AKT2 + AKT1
  • 宿主
    Rabbit
  • 特异性
    This product reacts with AKT1, AKT2 and AKT3.
  • 经测试应用
    适用于: ICC/IF, WB, IHC-P, Flow Cyt, IPmore details
  • 种属反应性
    与反应: Mouse, Human
    预测可用于: Rat, Cow
  • 免疫原

    Synthetic peptide within Human AKT1 aa 450 to the C-terminus (C terminal). The exact sequence is proprietary.
    Database link: P31749

  • 阳性对照
    • MCF7 cell lysate and prostate carcinoma tissue.
  • 常规说明

    A trial size is available to purchase for this antibody.

     

    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.

性能

应用

Our Abpromise guarantee covers the use of ab32505 in the following tested applications.

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

应用 Ab评论 说明
ICC/IF 1/100 - 1/250.
WB 1/2000 - 1/10000. Detects a band of approximately 59 kDa (predicted molecular weight: 56 kDa).
IHC-P 1/100.
Flow Cyt 1/20.

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

IP 1/100.

靶标

图片

  • ab32505 staining in SK-N-SH cells treated with alsterpaullone (ab141070), by ICC/IF. Decrease of AKT1 + AKT2 + AKT3 expression correlates with increased concentration of alsterpaullone, as described in literature.
    The cells were incubated at 37°C for 6h in media containing different concentrations of ab141070 (alsterpaullone) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab32505 (1/200 dilution was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.

  • Immunohistochemical analysis of paraffin-embedded prostate carcinoma using ab32505 at 1/100 dilution.
  • All lanes : Anti-AKT3 + AKT2 + AKT1 antibody [Y89] (ab32505) at 1/10000 dilution

    Lane 1 : 293T cell lysate transfected with GFP tagged AKT1
    Lanes 2 & 4 : 293T cell lysate transfected with empty vector
    Lane 3 : 293T cell lysate transfected with GFP tagged AKT3

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 56 kDa
    Observed band size: 82 kDa (why is the actual band size different from the predicted?)


    Exposure time: 8 seconds


    Blocking and diluting buffer and concentration: 5% NFDM/TBST

  • All lanes : Anti-AKT3 + AKT2 + AKT1 antibody [Y89] (ab32505) at 1/2000 dilution

    Lane 1 : 293T cell lysate transfected with GFP tagged AKT2
    Lane 2 : 293T cell lysate transfected with empty vector

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 56 kDa
    Observed band size: 82 kDa (why is the actual band size different from the predicted?)


    Exposure time: 5 seconds


    Blocking and diluting buffer and concentration: 5% NFDM/TBST

  • Anti-AKT3 + AKT2 + AKT1 antibody [Y89] (ab32505) at 1/10000 dilution + MCF-7 cell lysate

    Predicted band size: 56 kDa
    Observed band size: 59 kDa (why is the actual band size different from the predicted?)

  • Overlay histogram showing HeLa cells stained with ab32505 (red line). The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32505, 1/20 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a slightly decreased signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

文献

This product has been referenced in:
  • Qian G  et al. SeMet attenuates OTA-induced PCV2 replication promotion by inhibiting autophagy by activating the AKT/mTOR signaling pathway. Vet Res 49:15 (2018). Read more (PubMed: 29439710) »
  • Zhuo J  et al. Evaluation of type 2 diabetic mellitus animal models via interactions between insulin and mitogen-activated protein kinase signaling pathways induced by a high fat and sugar diet and streptozotocin. Mol Med Rep 17:5132-5142 (2018). Read more (PubMed: 29393432) »

See all 43 Publications for this product

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All the products you shortlisted seem to be suitable for your work, however, looking at the availability of those products, I would recommend the following one that are currently in stock:
Anti-AKT1 (N-term)ant...

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