Anti-AKT1 antibody [M1843] (ab108202)
![Western blot - Anti-AKT1 antibody [M1843] (ab108202)](https://www.abcam.com/ps/products/108/ab108202/Images/ab108202-484603-anti-akt1-antibody-m1843-western-blot-wildtype-mcf7-akt1-knockout-mcf7-daudi-a431.jpg "Western blot - Anti-AKT1 antibody [M1843] (ab108202)")
Key features and details
- Mouse monoclonal [M1843] to AKT1
- Suitable for: WB
- Knockout validated
- Reacts with: Human
- Isotype: IgG1
Overview
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Product name
Anti-AKT1 antibody [M1843]
See all AKT1 primary antibodies -
Description
Mouse monoclonal [M1843] to AKT1 -
Host species
Mouse -
Tested applications
Suitable for: WBmore details
Unsuitable for: Flow Cyt,ICC,IHC-P or IP -
Species reactivity
Reacts with: Human
Does not react with: Mouse, Rat -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: MCF7, Daudi, HeLa, A431 or A549 cell lysate.
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General notes
This mouse monoclonal to AKT1 has been knockout validated in Western blot. The expected band for AKT1 was observed in wild type cells and the band was not seen in knockout cells.
This product was manufactured under U.S. Patent No. 5,675,063.
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 50% Glycerol (glycerin, glycerine), 0.05% BSA, 49% PBS -
Concentration information loading... -
Purity
Tissue culture supernatant -
Clonality
Monoclonal -
Clone number
M1843 -
Isotype
IgG1 -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab108202 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 |
|---|---|---|
| WB |
1/1000 - 1/10000. Predicted molecular weight: 56 kDa.
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| Notes |
|---|
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WB
1/1000 - 1/10000. Predicted molecular weight: 56 kDa. |
Target
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Function
Plays a role as a key modulator of the AKT-mTOR signaling pathway controlling the tempo of the process of newborn neurons integration during adult neurogenesis, including correct neuron positioning, dendritic development and synapse formation (By similarity). General protein kinase capable of phosphorylating several known proteins. Phosphorylates TBC1D4. Signals downstream of phosphatidylinositol 3-kinase (PI(3)K) to mediate the effects of various growth factors such as platelet-derived growth factor (PDGF), epidermal growth factor (EGF), insulin and insulin-like growth factor I (IGF-I). Plays a role in glucose transport by mediating insulin-induced translocation of the GLUT4 glucose transporter to the cell surface. Mediates the antiapoptotic effects of IGF-I. Mediates insulin-stimulated protein synthesis by phosphorylating TSC2 at 'Ser-939' and 'Thr-1462', thereby activating mTORC1 signaling and leading to both phosphorylation of 4E-BP1 and in activation of RPS6KB1. Promotes glycogen synthesis by mediating the insulin-induced activation of glycogen synthase. The activated form can suppress FoxO gene transcription and promote cell cycle progression. Essential for the SPATA13-mediated regulation of cell migration and adhesion assembly and disassembly. -
Tissue specificity
Expressed in all human cell types so far analyzed. The Tyr-176 phosphorylated form shows a significant increase in expression in breast cancers during the progressive stages i.e. normal to hyperplasia (ADH), ductal carcinoma in situ (DCIS), invasive ductal carcinoma (IDC) and lymph node metastatic (LNMM) stages. -
Involvement in disease
Defects in AKT1 are a cause of susceptibility to breast cancer (BC) [MIM:114480]. A common malignancy originating from breast epithelial tissue. Breast neoplasms can be distinguished by their histologic pattern. Invasive ductal carcinoma is by far the most common type. Breast cancer is etiologically and genetically heterogeneous. Important genetic factors have been indicated by familial occurrence and bilateral involvement. Mutations at more than one locus can be involved in different families or even in the same case.
Defects in AKT1 are associated with colorectal cancer (CRC) [MIM:114500].
Defects in AKT1 are associated with susceptibility to ovarian cancer [MIM:604370]; also called susceptibility to familial breast-ovarian cancer type 1 (BROVCA1). -
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. The PH domain mediates interaction with TNK2 and Tyr-176 is also essential for this interaction.
The AGC-kinase C-terminal mediates interaction with THEM4. -
Post-translational
modificationsPhosphorylation on Thr-308, Ser-473 and Tyr-474 is required for full activity. Activated TNK2 phosphorylates it on Tyr-176 resulting in its binding to the anionic plasma membrane phospholipid PA. This phosphorylated form localizes to the cell membrane, where it is targeted by PDPK1 and PDPK2 for further phosphorylations on Thr-308 and Ser-473 leading to its activation. Ser-473 phosphorylation by mTORC2 favors Thr-308 phosphorylation by PDPK1. Ser-473 phosphorylation is enhanced by interaction with AGAP2 isoform 2 (PIKE-A). Ser-473 phosphorylation is enhanced in focal cortical dysplasias with Taylor-type balloon cells.
Ubiquitinated; undergoes both 'Lys-48'- and 'Lys-63'-linked polyubiquitination. TRAF6-induced 'Lys-63'-linked AKT1 ubiquitination is critical for phosphorylation and activation. When ubiquitinated, it translocates to the plasma membrane, where it becomes phosphorylated. 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. 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. - Information by UniProt
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Database links
- Entrez Gene: 207 Human
- Omim: 164730 Human
- SwissProt: P31749 Human
- Unigene: 525622 Human
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Alternative names
- AKT 1 antibody
- AKT antibody
- AKT1 antibody
see all
Images
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Western blot - Anti-AKT1 antibody [M1843] (ab108202)All lanes : Anti-AKT1 antibody [M1843] (ab108202) at 1/1000 dilution
Lane 1 : Wild-type MCF7 cell lysate
Lane 2 : AKT1 knockout MCF7 cell lysate
Lane 3 : Daudi cell lysate
Lane 4 : A431 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 56 kDa
Observed band size: 62 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-AKT1 antibody [M1843] staining at 1/1000 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab108202 was shown to bind specifically to AKT1. A band was observed at 62 kDa in wild-type MCF7 cell lysates with no signal observed at this size in AKT1 knockout cell line ab282330 (knockout cell lysate ab283003). To generate this image, wild-type and AKT1 knockout MCF7 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) at 1/20000 dilution.
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![Western blot - Anti-AKT1 antibody [M1843] (ab108202)](https://www.abcam.com/ps/products/108/ab108202/Images/ab108202-450623-anti-akt1-antibody-m1843-western-blot-hela-mcf7-daudi-lysates.jpg)
Western blot - Anti-AKT1 antibody [M1843] (ab108202)All lanes : Anti-AKT1 antibody [M1843] (ab108202) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : AKT1 knockout HeLa cell lysate
Lane 3 : MCF7 cell lysate
Lane 4 : Daudi cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 56 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted?Lanes 1- 4: Merged signal (red and green). Green - ab108202 observed at 60 kDa. Red - Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) observed at 37 kDa.
ab108202 was shown to react with AKT1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab264852 (knockout cell lysate ab256835) was used. Wild-type HeLa and AKT1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab108202 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye®800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye®680RD) preadsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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![Western blot - Anti-AKT1 antibody [M1843] (ab108202)](https://www.abcam.com/ps/products/108/ab108202/Images/ab108202-251554-anti-akt1-antibody-m1843-western-blot.jpg)
Western blot - Anti-AKT1 antibody [M1843] (ab108202)Lanes 1, 3 and 5: Wild-type HAP1 cell lysate (20 µg)
Lanes 2, 4 and 6: AKT1 knockout HAP1 cell lysate (20 µg)
Lanes 1 and 2: Green signal from target - ab108202 observed at 55 kDa
Lanes 3 and 4: Red signal from loading control - ab8227 observed at 42 kDa
Lanes 5 and 6: Merged (red and green) signal
ab108202 detected the expected band for AKT1 in wild-type cells and the band was not seen in knockout cells. Wild-type and AKT1 knockout samples were subjected to SDS-PAGE. ab108202 and ab8227 (loading control to beta actin) were both diluted at 1/1000 and incubated overnight at 4°C. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.
Datasheets and documents
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Datasheet download
References (3)
ab108202 has been referenced in 3 publications.
- Pillai-Kastoori L et al. Antibody validation for Western blot: By the user, for the user. J Biol Chem 295:926-939 (2020). PubMed: 31819006
- Cai Z et al. Immunoglobulin-like transcript 4 and human leukocyte antigen-G interaction promotes the progression of human colorectal cancer. Int J Oncol 54:1943-1954 (2019). PubMed: 30942436
- Chan JF et al. The celecoxib derivative kinase inhibitor AR-12 (OSU-03012) inhibits Zika virus via down-regulation of the PI3K/Akt pathway and protects Zika virus-infected A129 mice: A host-targeting treatment strategy. Antiviral Res 160:38-47 (2018). PubMed: 30326204
