Recombinant Anti-AKT1 (phospho S124) antibody [EPR17680] - BSA and Azide free (ab250676)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR17680] to AKT1 (phospho S124) - BSA and Azide free
- Suitable for: ELISA, Dot blot, IP, WB
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-AKT1 (phospho S124) antibody [EPR17680] - BSA and Azide free
See all AKT1 primary antibodies -
Description
Rabbit monoclonal [EPR17680] to AKT1 (phospho S124) - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: ELISA, Dot blot, IP, WBmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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General notes
ab250676 is the carrier-free version of ab183556.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR17680 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Assay kits
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Conjugation kits
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab250676 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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ELISA |
Use at an assay dependent concentration.
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Dot blot |
Use at an assay dependent concentration.
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IP |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration. Detects a band of approximately 56 kDa (predicted molecular weight: 56 kDa).
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Notes |
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ELISA
Use at an assay dependent concentration. |
Dot blot
Use at an assay dependent concentration. |
IP
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Detects a band of approximately 56 kDa (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
- Entrez Gene: 11651 Mouse
- Entrez Gene: 24185 Rat
- Omim: 164730 Human
- SwissProt: P31749 Human
- SwissProt: P31750 Mouse
- SwissProt: P47196 Rat
- Unigene: 525622 Human
see all -
Alternative names
- AKT 1 antibody
- AKT antibody
- AKT1 antibody
see all
Images
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All lanes : Anti-AKT1 (phospho S124) antibody [EPR17680] (ab183556) at 1/1000 dilution
Lane 1 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate, untreated
Lane 2 : MCF7 whole cell lysate, treated with Alkaline Phosphatase
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 kDa
Exposure time: 15 secondsThis data was developed using ab183556, the same antibody clone in a different buffer formulation.
Blocking and dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-AKT1 (phospho S124) antibody [EPR17680] (ab183556) at 1/10000 dilution
Lane 1 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate, untreated
Lane 2 : NIH/3T3 whole cell lysate, treated with Alkaline Phosphatase
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 kDa
Exposure time: 3 minutesThis data was developed using ab183556, the same antibody clone in a different buffer formulation.
Blocking and dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-AKT1 (phospho S124) antibody [EPR17680] (ab183556) at 1/10000 dilution
Lane 1 : C6 (Rat glial tumor cells) whole cell lysate, untreated
Lane 2 : C6 whole cell lysate, treated with Alkaline Phosphatase
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 kDa
Exposure time: 3 minutesThis data was developed using ab183556, the same antibody clone in a different buffer formulation.
Blocking and dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-AKT1 (phospho S124) antibody [EPR17680] (ab183556) at 1/1000 dilution
Lane 1 : Human fetal brain tissue lysate
Lane 2 : Human fetal kidney tissue lysate
Lane 3 : Mouse brain tissue lysate
Lane 4 : Rat brain tissue lysate
Lane 5 : Rat heart tissue lysate
Lane 6 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate
Lane 7 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell 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 kDa
Exposure time: 3 minutesThis data was developed using ab183556, the same antibody clone in a different buffer formulation.
Blocking and dilution buffer: 5% NFDM/TBST.
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This data was developed using ab183556, the same antibody clone in a different buffer formulation.
Immunoprecipitation of AKT1 from 1mg of MCF7 (Human breast adenocarcinoma cell line) whole cell lysate achieved using ab183556 at 1/100 dilution.
Lane 1: Input: 10µg of MCF7 whole cell lysate.
Lane 2: MCF7 whole cell lysate following IP with ab183556.
Lane 3: negative control: IP using Rabbit monoclonal IgG (ab172730) instead of ab183556 in MCF7 whole cell lysate.
Western blot was performed using ab183556 at 1/1000 dilution.
An Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1500 was used as secondary antibody. Blocking and dilution buffer: 5% NFDM/TBST.
Exposure time: 10 seconds.
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This data was developed using ab183556, the same antibody clone in a different buffer formulation.Dot blot analysis of AKT1 (phospho S124) labeled with ab183556 at 1/1000 dilution.
Lane 1: phospho peptide
Lane 2: non phospho peptide
Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 was used as secondary antibody.
Exposure time: 3 minutes.Blocking and Diluting buffer buffer and concentration: 5% NFDM/TBST -
This data was developed using ab183556, the same antibody clone in a different buffer formulation.ELISA image showing specificity of ab183556 to AKT1 (phospho S124) peptide only.Peptides concentration: 1000 ng/ml.ab183556 working dilution: 1/1700.Secondary antibody: Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/2500.
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.
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (1)
ab250676 has been referenced in 1 publication.
- Cheng Z et al. Upregulation of circRNA_100395 sponges miR-142-3p to inhibit gastric cancer progression by targeting the PI3K/AKT axis. Oncol Lett 21:419 (2021). PubMed: 33841580