Recombinant Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] (ab179463)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR16798] to AKT1 + AKT2 + AKT3
- Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra)
- Reacts with: Mouse, Rat, Human, Xenopus laevis, Xenopus tropicalis
Related conjugates and formulations
Overview
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Product name
Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798]
See all AKT1 + AKT2 + AKT3 primary antibodies -
Description
Rabbit monoclonal [EPR16798] to AKT1 + AKT2 + AKT3 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra)more details -
Species reactivity
Reacts with: Mouse, Rat, Human, Xenopus laevis, Xenopus tropicalis -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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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
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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
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
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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
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR16798 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
- Alexa Fluor® 488 Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] (ab200619)
- HRP Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] (ab200854)
- PE Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] (ab210454)
- Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] - Low endotoxin, Azide free (ab214167)
- Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] - BSA and Azide free (ab271930)
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Compatible Secondaries
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Conjugation kits
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab179463 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 |
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WB |
1/10000. Detects a band of approximately 56 kDa (predicted molecular weight: 56 kDa).
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IHC-P |
1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF |
1/100.
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IP |
1/100.
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Flow Cyt (Intra) |
1/330.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Notes |
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WB
1/10000. Detects a band of approximately 56 kDa (predicted molecular weight: 56 kDa). |
IHC-P
1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
1/100. |
IP
1/100. |
Flow Cyt (Intra)
1/330. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Target
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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
modificationsPhosphorylation 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
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Database links
- Entrez Gene: 10000 Human
- Entrez Gene: 207 Human
- Entrez Gene: 208 Human
- Entrez Gene: 11651 Mouse
- Entrez Gene: 11652 Mouse
- Entrez Gene: 23797 Mouse
- Entrez Gene: 24185 Rat
- Entrez Gene: 25233 Rat
see all -
Alternative names
- AKT antibody
- AKT1 antibody
- AKT1 kinase antibody
see all
Images
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All lanes : Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] (ab179463) at 1/10000 dilution
Lane 1 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysates
Lane 2 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysates
Lane 3 : Hep G2 (Human liver hepatocellular carcinoma) whole cell lysates
Lane 4 : A549 (Human lung carcinoma) whole cell lysates
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 56 kDa
Observed band size: 56 kDaBlocking/Dilution buffer: 5% NFDM/TBST.
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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. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] (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.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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All lanes : Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] (ab179463) at 1/1000 dilution
Lane 1 : Human fetal brain lysate
Lane 2 : Human fetal heart lysate
Lane 3 : Human fetal kidney lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 56 kDa
Observed band size: 56 kDaBlocking/Dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] (ab179463) at 1/1000 dilution
Lane 1 : Mouse brain lysate
Lane 2 : Mouse heart lysate
Lane 3 : Mouse kidney lysate
Lane 4 : Mouse spleen lysate
Lane 5 : Rat brain lysate
Lane 6 : Rat heart lysate
Lane 7 : Rat kidney lysate
Lane 8 : Rat spleen lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 56 kDa
Observed band size: 56 kDaBlocking/Dilution buffer: 5% NFDM/TBST.
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Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] (ab179463) at 1/10000 dilution + Xenopus muscle lysate at 10 µg
Secondary
Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 56 kDa
Observed band size: 56 kDaBlocking/Dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] (ab179463) at 1/10000 dilution
Lane 1 : AKT2 recombinant protein (HIS-tag): aa282-481
Lane 2 : AKT3 recombinant protein (HIS-tag) :aa351-479
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 56 kDa
Observed band size: 18,26 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] (ab179463)
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.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKT1 + AKT2 + AKT3 antibody [EPR16798] (ab179463)
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.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Intracellular 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.
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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.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (219)
ab179463 has been referenced in 219 publications.
- Liu F et al. Upregulation of circ_0000142 promotes multiple myeloma progression by adsorbing miR-610 and upregulating AKT3 expression. J Biochem 169:327-336 (2021). PubMed: 32970816
- Li Z et al. Kcnh2 mediates FAK/AKT-FOXO3A pathway to attenuate sepsis-induced cardiac dysfunction. Cell Prolif 54:e12962 (2021). PubMed: 33263944
- Niu L et al. Sericin inhibits MDA-MB-468 cell proliferation via the PI3K/Akt pathway in triple-negative breast cancer. Mol Med Rep 23:N/A (2021). PubMed: 33313947
- Li F et al. Combined transplantation of neural stem cells and bone marrow mesenchymal stem cells promotes neuronal cell survival to alleviate brain damage after cardiac arrest via microRNA-133b incorporated in extracellular vesicles. Aging (Albany NY) 13:262-278 (2021). PubMed: 33436530
- Zhou Y et al. The miR-204-5p/FOXC1/GDF7 axis regulates the osteogenic differentiation of human adipose-derived stem cells via the AKT and p38 signalling pathways. Stem Cell Res Ther 12:64 (2021). PubMed: 33461605