Recombinant Anti-PRAS40 antibody [EPR6402] (ab133584)
![Western blot - Anti-PRAS40 antibody [EPR6402] (ab133584)](https://www.abcam.com/ps/products/133/ab133584/Images/ab133584-319569-anti-pras40-antibody-epr6402-western-blot.jpg "Western blot - Anti-PRAS40 antibody [EPR6402] (ab133584)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR6402] to PRAS40
- Suitable for: WB
- Knockout validated
- Reacts with: Mouse, Rat, Human
Overview
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Product name
Anti-PRAS40 antibody [EPR6402]
See all PRAS40 primary antibodies -
Description
Rabbit monoclonal [EPR6402] to PRAS40 -
Host species
Rabbit -
Tested applications
Suitable for: WBmore details
Unsuitable for: Flow Cyt,ICC or IHC-P -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide within Human PRAS40 aa 150-250 (internal sequence). The exact sequence is proprietary.
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Positive control
- MCF7, L6, C6, RAW 264.7, PC12, NIH 3T3, 293T and HeLa whole cell lysate (ab150035)
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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.
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 -20°C. Stable for 12 months at -20°C. -
Dissociation constant (KD)
KD = 1.43 x 10 -11 M Learn more about KD -
Storage buffer
pH: 7.20
Preservative: 0.05% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant -
Concentration information loading... -
Purity
Tissue culture supernatant -
Clonality
Monoclonal -
Clone number
EPR6402 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
Applications
Our Abpromise guarantee covers the use of ab133584 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. Detects a band of approximately 40 kDa (predicted molecular weight: 27 kDa). |
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
- Entrez Gene: 292887 Rat
- 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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Western blot - Anti-PRAS40 antibody [EPR6402] (ab133584)All lanes : Anti-PRAS40 antibody [EPR6402] (ab133584) 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 - ab133584 observed at 40 kDa. Red - loading control, ab130007, observed at 125 kDa.
ab133584 was shown to specifically react with PRAS40 in wild-type HAP1 cells as signal was lost in AKT1S1 (PRAS40) knockout cells. Wild-type and AKT1S1 (PRAS40) knockout samples were subjected to SDS-PAGE. Ab133584 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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![Western blot - Anti-PRAS40 antibody [EPR6402] (ab133584)](https://www.abcam.com/ps/products/133/ab133584/Images/ab133584-147391-anti-pras40-antibody-epr6402-western-blot.jpg)
Western blot - Anti-PRAS40 antibody [EPR6402] (ab133584)All lanes : Anti-PRAS40 antibody [EPR6402] (ab133584) at 1/1000 dilution
Lane 1 : MCF7 cell lysate
Lane 2 : L6 cell lysate
Lane 3 : C6 cell lysate
Lane 4 : RAW 264.7 cell lysate
Lane 5 : PC12 cell lysate
Lane 6 : NIH 3T3 cell lysate
Lane 7 : 293T cell lysate
Lane 8 : HeLa cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP labelled Goat anti-Rabbit IgG at 1/2000 dilution
Predicted band size: 27 kDa -
![OI-RD Scanning - Anti-PRAS40 antibody [EPR6402] (ab133584)](https://www.abcam.com/ps/products/133/ab133584/Images/ab133584-147385-anti-pras40-antibody-epr6402.jpg)
OI-RD Scanning - Anti-PRAS40 antibody [EPR6402] (ab133584) -
![Anti-PRAS40 antibody [EPR6402] (ab133584)](https://www.abcam.com/ps/products/133/ab133584/Images/ab133584-9-benefits-of-recombinant-antibodies.png)
Anti-PRAS40 antibody [EPR6402] (ab133584)
Datasheets and documents
References (0)
ab133584 has not yet been referenced specifically in any publications.
