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

製品の概要

  • 製品名Anti-AKT3 + AKT2 + AKT1 antibody [Y89]
  • 製品の詳細
    Rabbit monoclonal [Y89] to AKT3 + AKT2 + AKT1
  • 特異性This product reacts with AKT1, AKT2 and AKT3.
  • アプリケーション適用あり: ICC/IF, WB, IHC-P, Flow Cyt, IPmore details
  • 種交差性
    交差種: Mouse, Human
    交差が予測される動物種: Rat, Cow
  • 免疫原

    corresponding to Human AKT1 aa 450 to the C-terminus (C terminal).
    Database link: P31749

  • ポジティブ・コントロール
    • MCF7 cell lysate and prostate carcinoma tissue.
  • 特記事項

    This product is a recombinant rabbit monoclonal antibody.

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

    A trial size is available to purchase for this antibody.

    Alternative versions available:

    Anti-AKT1 antibody (Alexa Fluor® 488) [Y89] (ab204252)
    Anti-AKT1 antibody (Alexa Fluor® 647) [Y89] (ab204285)

     

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
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.

ターゲット情報

Anti-AKT3 + AKT2 + AKT1 antibody [Y89] 画像

  • 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
    Lane 2 : 293T cell lysate transfected with empty vector
    Lane 3 : 293T cell lysate transfected with GFP tagged AKT3
    Lane 4 : 293T cell lysate transfected with empty vector

    Lysates/proteins at 10 µg per lane.

    Secondary
    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
    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.

Anti-AKT3 + AKT2 + AKT1 antibody [Y89] (ab32505) 使用論文

This product has been referenced in:
  • Chen Y  et al. FGFR antagonist induces protective autophagy in FGFR1-amplified breast cancer cell. Biochem Biophys Res Commun N/A:N/A (2016). WB ; Human . Read more (PubMed: 26993162) »
  • Liang D  et al. Therapeutic efficacy of apelin on transplanted mesenchymal stem cells in hindlimb ischemic mice via regulation of autophagy. Sci Rep 6:21914 (2016). WB ; Mouse . Read more (PubMed: 26902855) »

See all 26 Publications for this product

Product Wall

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (Bile duct ligation in Liver)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citrate buffer ph6 20 min at 97ºC in a PT link
Permeabilization No
Specification Bile duct ligation in Liver
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative Formaldehyde
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投稿 Sep 24 2015

Application Western blot
Loading amount 40 µg
Gel Running Conditions Reduced Denaturing
Sample Mouse Cell lysate - whole cell (Cardiomyocytes)
Specification Cardiomyocytes
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
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投稿 Feb 11 2015

Application Western blot
Loading amount 25 µg
Gel Running Conditions Reduced Denaturing (4–15%)
Sample Mouse Tissue lysate - whole (Lung)
Specification Lung
Blocking step BSA as blocking agent for 15 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
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投稿 Jul 09 2014

Thank you for contacting us.

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...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (Glioblastoma Initiating Cells (Primary cells))
Loading amount 30 µg
Specification Glioblastoma Initiating Cells (Primary cells)
Gel Running Conditions Reduced Denaturing (12.5)
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 20°C
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投稿 Nov 16 2011

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"