Recombinant Anti-PRAS40 antibody [EPR6263(2)] (ab151719)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR6263(2)] to PRAS40
- Suitable for: WB, IHC-P, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Mouse, Human
Related conjugates and formulations
Overview
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Product name
Anti-PRAS40 antibody [EPR6263(2)]
See all PRAS40 primary antibodies -
Description
Rabbit monoclonal [EPR6263(2)] to PRAS40 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, Flow Cyt (Intra)more details
Unsuitable for: ICC/IF -
Species reactivity
Reacts with: Mouse, Human -
Immunogen
Synthetic peptide within Human PRAS40 aa 150-250. The exact sequence is proprietary.
Database link: Q96B36 -
Positive control
- 293T, SHSY5Y, HeLa and NIH 3T3 cell lysates; Human liver tissue; 293T cells.
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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.
Rat: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20ºC. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.5% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR6263(2) -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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Positive Controls
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab151719 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 |
1/1000 - 1/10000. Detects a band of approximately 40 kDa (predicted molecular weight: 27 kDa).
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IHC-P |
1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Flow Cyt (Intra) |
1/10 - 1/100.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Notes |
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WB
1/1000 - 1/10000. Detects a band of approximately 40 kDa (predicted molecular weight: 27 kDa). |
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
Flow Cyt (Intra)
1/10 - 1/100. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Target
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Function
Subunit of mTORC1, which regulates cell growth and survival in response to nutrient and hormonal signals. mTORC1 is activated in response to growth factors or amino-acids. Growth factor-stimulated mTORC1 activation involves a AKT1-mediated phosphorylation of TSC1-TSC2, which leads to the activation of the RHEB GTPase that potently activates the protein kinase activity of mTORC1. Amino-acid-signaling to mTORC1 requires its relocalization to the lysosomes mediated by the Ragulator complex and the Rag GTPases. Activated mTORC1 up-regulates protein synthesis by phosphorylating key regulators of mRNA translation and ribosome synthesis. mTORC1 phosphorylates EIF4EBP1 and releases it from inhibiting the elongation initiation factor 4E (eiF4E). mTORC1 phosphorylates and activates S6K1 at 'Thr-389', which then promotes protein synthesis by phosphorylating PDCD4 and targeting it for degradation. Within mTORC1, AKT1S1 negatively regulates mTOR activity in a manner that is dependent on its phosphorylation state and binding to 14-3-3 proteins. Inhibits RHEB-GTP-dependent mTORC1 activation. Substrate for AKT1 phosphorylation, but can also be activated by AKT1-independent mechanisms. May also play a role in nerve growth factor-mediated neuroprotection. -
Tissue specificity
Widely expressed with highest levels of expression in liver and heart. Expressed at higher levels in cancer cell lines (e.g. A549 and HeLa) than in normal cell lines (e.g. HEK293). -
Post-translational
modificationsPhosphorylated by AKT1. Phosphorylation relieves inhibitory function on mTORC1. -
Cellular localization
Cytoplasm > cytosol. Found in the cytosolic fraction of the brain. - Information by UniProt
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Database links
- Entrez Gene: 84335 Human
- Entrez Gene: 67605 Mouse
- Omim: 610221 Human
- SwissProt: Q96B36 Human
- SwissProt: Q9D1F4 Mouse
- Unigene: 515542 Human
- Unigene: 148007 Mouse
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Alternative names
- 40 kDa proline rich AKT substrate antibody
- 40 kDa proline-rich AKT substrate antibody
- AKT1 S1 antibody
see all
Images
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All lanes : Anti-PRAS40 antibody [EPR6263(2)] (ab151719) at 1/1000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : AKT1S1 knockout HAP1 whole cell lysate
Lane 3 : HeLa whole cell lysate
Lane 4 : MCF-7 whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 27 kDa
Observed band size: 40 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab151719 observed at 27 kDa. Red - loading control, ab130007, observed at 124 kDa.
ab151719 was shown to recognize PRAS40 in wild-type HAP1 cells as signal was lost at the expected MW in AKT1S1 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and AKT1S1 knockout samples were subjected to SDS-PAGE. Ab151719 and ab130007 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling PRAS40 with ab151719 at 1/50 dilution.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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All lanes : Anti-PRAS40 antibody [EPR6263(2)] (ab151719) at 1/1000 dilution
Lane 1 : 293T cell lysate
Lane 2 : SHSY5Y cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : NIH 3T3 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat anti-rabbit HRP at 1/2000 dilution
Predicted band size: 27 kDa
Observed band size: 40 kDa why is the actual band size different from the predicted? -
Intracellular flow cytometric analysis of permeabilized 293T cells labeling PRAS40 with ab151719 at 1/10 dilution (red), compared to a nonspecific control antibody (green).
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (5)
ab151719 has been referenced in 5 publications.
- Huang Y et al. Identification and validation of a cigarette smoke-related five-gene signature as a prognostic biomarker in kidney renal clear cell carcinoma. Sci Rep 12:2189 (2022). PubMed: 35140327
- Wang C et al. Long Noncoding RNA LINC01134 Promotes Hepatocellular Carcinoma Metastasis via Activating AKT1S1 and NF-?B Signaling. Front Cell Dev Biol 8:429 (2020). PubMed: 32656205
- Wang W et al. PTEN inhibitor VO-OHpic suppresses TSC2-/- MEFs proliferation by excessively inhibiting autophagy via the PTEN/PRAS40 pathway. Exp Ther Med 19:3565-3570 (2020). PubMed: 32346419
- Zhao S et al. The Apoptosis Regulator 14-3-3? and Its Potential as a Therapeutic Target in Pituitary Oncocytoma. Front Endocrinol (Lausanne) 10:797 (2019). PubMed: 31849836
- Xu Q et al. Inhibition of PTP1B blocks pancreatic cancer progression by targeting the PKM2/AMPK/mTOC1 pathway. Cell Death Dis 10:874 (2019). PubMed: 31745071