Anti-AKT3 + AKT2 + AKT1 antibody [Y89] (ab32505)
- Datasheet
- Specific References (43)
- Protocols
Overview
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Product nameAnti-AKT3 + AKT2 + AKT1 antibody [Y89]
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DescriptionRabbit monoclonal [Y89] to AKT3 + AKT2 + AKT1
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Host speciesRabbit
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SpecificityThis product reacts with AKT1, AKT2 and AKT3.
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Tested applicationsSuitable for: ICC/IF, WB, IHC-P, Flow Cyt, IPmore details
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Species reactivityReacts with: Mouse, Human
Predicted to work with: Rat, Cow
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Immunogen
Synthetic peptide within Human AKT1 aa 450 to the C-terminus (C terminal). The exact sequence is proprietary.
Database link: P31749 -
Positive control
- MCF7 cell lysate and prostate carcinoma tissue.
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General notes
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.
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
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Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 49% PBS, 50% Glycerol, 0.05% BSA -
Concentration information loading... -
PurityProtein A purified
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ClonalityMonoclonal
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Clone numberY89
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IsotypeIgG
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Research areas
Associated products
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Alternative Versions
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Assay kits
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Compatible Secondaries
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Positive Controls
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Related Products
Applications
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.
| Application | Abreviews | Notes |
|---|---|---|
| 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. |
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| IP | 1/100. |
Target
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Cellular localizationAKT3: Cytoplasm. Membrane. Membrane-associated after cell stimulation leading to its translocation. AKT1: Cytoplasm. Nucleus. Cell membrane. Nucleus after activation by integrin-linked protein kinase 1 (ILK1). Nuclear translocation is enhanced by interaction with TCL1A. Phosphorylation on Tyr-176 by TNK2 results in its localization to the cell membrane where it is targeted for further phosphorylations on Thr-308 and Ser-473 leading to its activation and the activated form translocates to the nucleus.
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Database links
- Entrez Gene: 100137872 Cow
- Entrez Gene: 280991 Cow
- Entrez Gene: 534923 Cow
- Entrez Gene: 10000 Human
- Entrez Gene: 207 Human
- Entrez Gene: 208 Human
- Entrez Gene: 11651 Mouse
- Entrez Gene: 11652 Mouse
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Images
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![Immunocytochemistry/ Immunofluorescence - Anti-AKT3 + AKT2 + AKT1 antibody [Y89] (ab32505)](https://www.abcam.com/ps/products/32/ab32505/Images/ab32505-4-ab141070smol.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-AKT3 + AKT2 + AKT1 antibody [Y89] (ab32505)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. -
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKT3 + AKT2 + AKT1 antibody [Y89] (ab32505)](https://www.abcam.com/ps/products/32/ab32505/Images/ab32505ihc.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKT3 + AKT2 + AKT1 antibody [Y89] (ab32505)Immunohistochemical analysis of paraffin-embedded prostate carcinoma using ab32505 at 1/100 dilution. -
![Western blot - Anti-AKT3 + AKT2 + AKT1 antibody [Y89] (ab32505)](https://www.abcam.com/ps/products/32/ab32505/Images/ab32505-271510-4.jpg)
Western blot - Anti-AKT3 + AKT2 + AKT1 antibody [Y89] (ab32505)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 secondsBlocking and diluting buffer and concentration: 5% NFDM/TBST
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![Western blot - Anti-AKT3 + AKT2 + AKT1 antibody [Y89] (ab32505)](https://www.abcam.com/ps/products/32/ab32505/Images/ab32505-271511-6.jpg)
Western blot - Anti-AKT3 + AKT2 + AKT1 antibody [Y89] (ab32505)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 secondsBlocking and diluting buffer and concentration: 5% NFDM/TBST
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![Western blot - Anti-AKT3 + AKT2 + AKT1 antibody [Y89] (ab32505)](https://www.abcam.com/ps/products/32/ab32505/Images/ab32505.gif)
Western blot - Anti-AKT3 + AKT2 + AKT1 antibody [Y89] (ab32505)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?) -
![Flow Cytometry - Anti-AKT3 + AKT2 + AKT1 antibody [Y89] (ab32505)](https://www.abcam.com/ps/products/32/ab32505/Images/AKT1-Primary-antibodies-ab32505-1.jpg)
Flow Cytometry - Anti-AKT3 + AKT2 + AKT1 antibody [Y89] (ab32505)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.
References
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) »
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