Anti-AKT1 antibody (ab235958)
Key features and details
- Rabbit polyclonal to AKT1
- Suitable for: ChIP, IP, IHC-P, WB, ICC/IF
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-AKT1 antibody
See all AKT1 primary antibodies -
Description
Rabbit polyclonal to AKT1 -
Host species
Rabbit -
Tested applications
Suitable for: ChIP, IP, IHC-P, WB, ICC/IFmore details -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rat, Cow, Xenopus laevis
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Immunogen
Recombinant full length protein corresponding to Human AKT1 aa 1-480.
Sequence:MSDVAIVKEGWLHKRGEYIKTWRPRYFLLKNDGTFIGYKERPQDVDQREA PLNNFSVAQCQLMKTERPRPNTFIIRCLQWTTVIERTFHVETPEEREEWT TAIQTVADGLKKQEEEEMDFRSGSPSDNSGAEEMEVSLAKPKHRVTMNEF EYLKLLGKGTFGKVILVKEKATGRYYAMKILKKEVIVAKDEVAHTLTENR VLQNSRHPFLTALKYSFQTHDRLCFVMEYANGGELFFHLSRERVFSEDRA RFYGAEIVSALDYLHSEKNVVYRDLKLENLMLDKDGHIKITDFGLCKEGI KDGATMKTFCGTPEYLAPEVLEDNDYGRAVDWWGLGVVMYEMMCGRLPFY NQDHEKLFELILMEEIRFPRTLGPEAKSLLSGLLKKDPKQRLGGGSEDAK EIMQHRFFAGIVWQHVYEKKLSPPFKPQVTSETDTRYFDEEFTAQMITIT PPDQDDSMECVDSERRPHFPQFSYSASGTA
Database link: P31749-1 -
Positive control
- WB: HeLa, A549, HepG2 and MCF7 whole cell lysate. IHC-P: Human prostate, tonsil, cervical cancer and brain tissue. ICC/IF: HeLa cells. IP: HepG2 whole cell lysate.
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General notes
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
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, as well as customer reviews and Q&As.
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
pH: 7.40
Constituents: 50% Glycerol, PBS, 0.03% Proclin 300 -
Concentration information loading... -
Purity
Protein G purified -
Purification notes
Purity >95% -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
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Related Products
Applications
Our Abpromise guarantee covers the use of ab235958 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 |
|---|---|---|
| ChIP | Use 4µg for 106 cells. | |
| IP | 1/200 - 1/2000. | |
| IHC-P | 1/20 - 1/200. | |
| WB | 1/500 - 1/5000. | |
| ICC/IF | 1/100 - 1/500. |
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: 280991 Cow
- Entrez Gene: 207 Human
- Entrez Gene: 11651 Mouse
- Entrez Gene: 24185 Rat
- Entrez Gene: 399170 Xenopus laevis
- Omim: 164730 Human
- SwissProt: Q01314 Cow
- SwissProt: P31749 Human
see all -
Alternative names
- AKT 1 antibody
- AKT antibody
- AKT1 antibody
see all
Images
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Western blot - Anti-AKT1 antibody (ab235958)All lanes : Anti-AKT1 antibody (ab235958) at 1/500 dilution
Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : A549 (human lung carcinoma cell line) whole cell lysate
Lane 3 : HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate
Lane 4 : MCF7 (human breast adenocarcinoma cell line) whole cell lysate
Secondary
All lanes : Goat polyclonal to rabbit IgG at 1/50000 dilution -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKT1 antibody (ab235958)Paraffin-embedded human cervical cancer tissue stained for AKT1 using ab235958 at 1/100 dilution in immunohistochemical analysis.
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ChIP - Anti-AKT1 antibody (ab235958)Chromatin Immunoprecipitation HeLa (Human epithelial cell line from cervix adenocarcinoma) (1.1x106) were cross-linked with formaldehyde, sonicated and immunoprecipitated with 4 µg of ab235958 or a control normal rabbit IgG. The resulting ChIP DNA was quantified tissue using real-time PCR with primers against the exon-1 of Egr1 promoter.
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Immunocytochemistry/ Immunofluorescence - Anti-AKT1 antibody (ab235958)HeLa (Human epithelial cell line from cervix adenocarcinoma) cells stained for AKT1 (Green) using ab235958 at a 1/100 dilution in ICC/IF. Secondary used is an Alexa-Fluor®488-conjugated Goat Anti-Rabbit IgG (H+L). Counterstained with DAPI (Blue).
The cells were fixed in 4% formaldehyde, permeabilized using 0.2% Triton X-100 and blocked in 10% normal goat serum. The cells were then incubated with the primary antibody overnight at 4°C.
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Immunoprecipitation - Anti-AKT1 antibody (ab235958)AKT1 was immunoprecipitated from 500 μg of HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate. For western blotting, an HRP-conjugated Protein G antibody was used as the secondary antibody at 1/2000 dilution.
Lane 1: Rabbit control IgG instead of ab235958.
Lane 2: ab235958 IP in HepG2 whole cell lysate.
Lane 3: HepG2 whole cell lysate (input). -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKT1 antibody (ab235958)Paraffin-embedded human prostate tissue stained for AKT1 using ab235958 at 1/100 dilution in immunohistochemical analysis.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKT1 antibody (ab235958)Paraffin-embedded human brain tissue stained for AKT1 using ab235958 at 1/200 dilution in immunohistochemical analysis.
After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30 minutes at RT. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a biotinylated secondary antibody and visualized tissue using an HRP conjugated SP system.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKT1 antibody (ab235958)Paraffin-embedded human tonsil tissue stained for AKT1 using ab235958 at 1/100 dilution in immunohistochemical analysis.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
References (10)
ab235958 has been referenced in 10 publications.
- Zhou X et al. Long noncoding RNA BSN-AS2 induced by E2F1 promotes spinal osteosarcoma progression by targeting miR-654-3p/SYTL2 axis. Cancer Cell Int 20:133 (2020). PubMed: 32351327
- Wang B et al. Upregulation of contactin-1 expression promotes prostate cancer progression. Oncol Lett 19:1611-1618 (2020). PubMed: 32002038
- Wei F et al. Plasma endothelial cells-derived extracellular vesicles promote wound healing in diabetes through YAP and the PI3K/Akt/mTOR pathway. Aging (Albany NY) 12:12002-12018 (2020). PubMed: 32570219
- Xie T et al. microRNA-582 Potentiates Liver and Lung Metastasis of Gastric Carcinoma Cells Through the FOXO3-Mediated PI3K/Akt/Snail Pathway. Cancer Manag Res 12:5201-5212 (2020). PubMed: 32636681
- Luo Y et al. Effects of MiR-107 on The Chemo-drug Sensitivity of Breast Cancer Cells. Open Med (Wars) 14:59-65 (2019). PubMed: 31346547
- Sang B et al. Ras-AKT signaling represses the phosphorylation of histone H1.5 at threonine 10 via GSK3 to promote the progression of glioma. Artif Cells Nanomed Biotechnol 47:2882-2890 (2019). PubMed: 31307224
- Li C et al. HIF-1a/VEGF signaling-mediated epithelial-mesenchymal transition and angiogenesis is critically involved in anti-metastasis effect of luteolin in melanoma cells. Phytother Res 33:798-807 (2019). PubMed: 30653763
- Huo X et al. Clinical and Expression Significance of AKT1 by Co-expression Network Analysis in Endometrial Cancer. Front Oncol 9:1147 (2019). PubMed: 31781484
- Tian Y et al. CXCL12 induces migration of oligodendrocyte precursor cells through the CXCR4-activated MEK/ERK and PI3K/AKT pathways. Mol Med Rep 18:4374-4380 (2018). PubMed: 30221695
- Liu F et al. Overexpression of SENP1 reduces the stemness capacity of osteosarcoma stem cells and increases their sensitivity to HSVtk/GCV. Int J Oncol 53:2010-2020 (2018). PubMed: 30226577
