Recombinant Anti-AKT1 + AKT2 + AKT3 antibody [EPR17671] (ab185633)
- Datasheet
- References (4)
- Protocols
Overview
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Product name
Anti-AKT1 + AKT2 + AKT3 antibody [EPR17671]
See all AKT1 + AKT2 + AKT3 primary antibodies -
Description
Rabbit monoclonal [EPR17671] to AKT1 + AKT2 + AKT3 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt, IP, WB, IHC-P, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment within Human AKT1 + AKT2 + AKT3 aa 350 to the C-terminus. The exact sequence is proprietary.
Database link: Q9Y243 -
Positive control
- WB: AKT3 recombinant protein fragment (His-Tag®): aa351-479; AKT2 recombinant protein fragment (His-Tag®): aa282-481; AKT1 recombinant protein fragment (His-Tag®): aa281-480; A549 whole cell lysate; Human fetal brain and fetal kidney lysates; Mouse brain lysate; Rat brain and heart lysates. IHC-P: Human cerebral cortex, Human adenocarcinoma of colon, mouse cerebral cortex and rat kidney tissues. ICC/IF: HeLa cells. Flow Cytometry: A549 cells. IP: A549 whole cell extract.
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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... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR17671 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
Applications
Our Abpromise guarantee covers the use of ab185633 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 |
|---|---|---|
| Flow Cyt | 1/50. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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| IP | 1/50. | |
| WB | 1/2000. Detects a band of approximately 56 kDa (predicted molecular weight: 56 kDa). | |
| IHC-P | 1/400. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. | |
| ICC/IF | 1/500. |
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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![Western blot - Anti-AKT1 + AKT2 + AKT3 antibody [EPR17671] (ab185633)](https://www.abcam.com/ps/products/185/ab185633/Images/ab185633-237228-anti-akt1-2-3-antibody-epr17671-western-blot.jpg)
Western blot - Anti-AKT1 + AKT2 + AKT3 antibody [EPR17671] (ab185633)All lanes : Anti-AKT1 + AKT2 + AKT3 antibody [EPR17671] (ab185633) at 1/5000 dilution
Lane 1 : AKT3 recombinant protein fragment (His-Tag®): aa351-479
Lane 2 : AKT2 recombinant protein fragment (His-Tag®): aa282-481
Lane 3 : AKT1 recombinant protein fragment (His-Tag®): aa281-480
Lysates/proteins at 0.01 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 kDaBlocking/Dilution buffer: 5% NFDM/TBST.
The bottom panel is probed with an anti-His tag antibody.
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![Western blot - Anti-AKT1 + AKT2 + AKT3 antibody [EPR17671] (ab185633)](https://www.abcam.com/ps/products/185/ab185633/Images/ab185633-237229-anti-akt1-2-3-antibody-epr17671-western-blot.jpg)
Western blot - Anti-AKT1 + AKT2 + AKT3 antibody [EPR17671] (ab185633)Anti-AKT1 + AKT2 + AKT3 antibody [EPR17671] (ab185633) at 1/2000 dilution + A549 (Human lung carcinoma) whole cell lysates at 20 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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![Western blot - Anti-AKT1 + AKT2 + AKT3 antibody [EPR17671] (ab185633)](https://www.abcam.com/ps/products/185/ab185633/Images/ab185633-237230-anti-akt1-2-3-antibody-epr17671-western-blot.jpg)
Western blot - Anti-AKT1 + AKT2 + AKT3 antibody [EPR17671] (ab185633)All lanes : Anti-AKT1 + AKT2 + AKT3 antibody [EPR17671] (ab185633) at 1/2000 dilution
Lane 1 : Human fetal brain lysates
Lane 2 : Human fetal kidney lysates
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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![Western blot - Anti-AKT1 + AKT2 + AKT3 antibody [EPR17671] (ab185633)](https://www.abcam.com/ps/products/185/ab185633/Images/ab185633-237231-anti-akt1-2-3-antibody-epr17671-western-blot.jpg)
Western blot - Anti-AKT1 + AKT2 + AKT3 antibody [EPR17671] (ab185633)All lanes : Anti-AKT1 + AKT2 + AKT3 antibody [EPR17671] (ab185633) at 1/2000 dilution
Lane 1 : Mouse brain lysates
Lane 2 : Rat brain lysates
Lane 3 : Rat heart lysates
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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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKT1 + AKT2 + AKT3 antibody [EPR17671] (ab185633)](https://www.abcam.com/ps/products/185/ab185633/Images/ab185633-237223-anti-akt1-2-3-antibody-epr17671-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKT1 + AKT2 + AKT3 antibody [EPR17671] (ab185633)Immunohistochemical analysis of paraffin-embedded Human cerebral cortex tissue labeling AKT1 + AKT2 + AKT3 with ab185633 at 1/400 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear and weak cytoplasmic staining on neurons of the Human cerebral cortex is observed. Counter stained with Hematoxylin.
Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
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 [EPR17671] (ab185633)](https://www.abcam.com/ps/products/185/ab185633/Images/ab185633-237224-anti-akt1-2-3-antibody-epr17671-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKT1 + AKT2 + AKT3 antibody [EPR17671] (ab185633)Immunohistochemical analysis of paraffin-embedded Human adenocarcinoma of colon tissue labeling AKT1 + AKT2 + AKT3 with ab185633 at 1/400 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear and weak cytoplasmic staining on Human adenocarcinoma of colon is observed. Counter stained with Hematoxylin.
Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (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 + AKT3 antibody [EPR17671] (ab185633)](https://www.abcam.com/ps/products/185/ab185633/Images/ab185633-237225-anti-akt1-2-3-antibody-epr17671-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKT1 + AKT2 + AKT3 antibody [EPR17671] (ab185633)Immunohistochemical analysis of paraffin-embedded Mouse cerebral cortex tissue labeling AKT1 + AKT2 + AKT3 with ab185633 at 1/400 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear and weak cytoplasmic staining on neurons of the mouse cerebral cortex is observed. Counter stained with Hematoxylin.
Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (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 + AKT3 antibody [EPR17671] (ab185633)](https://www.abcam.com/ps/products/185/ab185633/Images/ab185633-237226-anti-akt1-2-3-antibody-epr17671-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKT1 + AKT2 + AKT3 antibody [EPR17671] (ab185633)Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling AKT1 + AKT2 + AKT3 with ab185633 at 1/400 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear and weak cytoplasmic staining on rat kidney is observed. Counter stained with Hematoxylin.
Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
-
![Immunoprecipitation - Anti-AKT1 + AKT2 + AKT3 antibody [EPR17671] (ab185633)](https://www.abcam.com/ps/products/185/ab185633/Images/ab185633-237227-anti-akt1-2-3-antibody-epr17671-immunoprecipitation.jpg)
Immunoprecipitation - Anti-AKT1 + AKT2 + AKT3 antibody [EPR17671] (ab185633)AKT1 + AKT2 + AKT3 was immunoprecipitated from 1mg of A549 (Human lung carcinoma) whole cell extract with ab185633 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab185633 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: A549 whole cell extract 10 µg (Input). Lane 2: ab185633 IP in A549 whole cell extract. Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab185633 in A549 whole cell extract.
Blocking and dilution buffer and concentration: 5% NFDM/TBST. -
![Flow Cytometry - Anti-AKT1 + AKT2 + AKT3 antibody [EPR17671] (ab185633)](https://www.abcam.com/ps/products/185/ab185633/Images/ab185633-237221-anti-akt1-2-3-antibody-epr17671-flow-cytometry.jpg)
Flow Cytometry - Anti-AKT1 + AKT2 + AKT3 antibody [EPR17671] (ab185633)Flow cytometric analysis of 2% paraformaldehyde-fixed A549 (Human lung carcinoma) cells labeling AKT1 + AKT2 + AKT3 with ab185633 at 1/50 dilution (red) compared with a rabbit monoclonal IgG isotype control (black) and an 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.
Protocols
Datasheets and documents
References
This product has been referenced in:
- Zhang X et al. Emodin protects H9c2 cells from hypoxia-induced injury by up-regulating miR-138 expression. Braz J Med Biol Res 52:e7994 (2019). Read more (PubMed: 30810622) »
- Wang M et al. Protective effects of Myrica rubra flavonoids against hypoxia/reoxygenation-induced cardiomyocyte injury via the regulation of the PI3K/Akt/GSK3ß pathway. Int J Mol Med 43:2133-2143 (2019). Read more (PubMed: 30864694) »
