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Purified anti-PRAS40/AKT1S1 Antibody

Pricing & Availability
Clone
W22030A (See other available formats)
Regulatory Status
RUO
Other Names
AKT1S1, AKT1 Substrate 1, PRAS40, 40 KDa Proline-Rich AKT Substrate, AKT1 Substrate 1, Proline-Rich AKT1 Substrate 1, MGC2865
Isotype
Rat IgG2a, κ
Ave. Rating
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Product Citations
publications
A. W22030A_PURE_PRAS40_WB_072224
Whole cell extracts (15 µg total protein per lane) from the indicated cell lines were resolved on a 4-12% Bis-Tris gel, transferred to a PVDF membrane, and probed with Purified anti-PRAS40/AKT1S1 (clone W22030A) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP Goat anti-rat IgG (Cat. No. 405405) at a 1:5000 dilution. Direct-Blot™ HRP anti-β-actin (Cat. No. 664804) was used as a loading control at a 1:25000 dilution. Western-Ready™ ECL Substrate Premium Kit (Cat. No. 426319) was used as a detection agent. Lane M: Molecular weight marker.
  • A. W22030A_PURE_PRAS40_WB_072224
    Whole cell extracts (15 µg total protein per lane) from the indicated cell lines were resolved on a 4-12% Bis-Tris gel, transferred to a PVDF membrane, and probed with Purified anti-PRAS40/AKT1S1 (clone W22030A) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP Goat anti-rat IgG (Cat. No. 405405) at a 1:5000 dilution. Direct-Blot™ HRP anti-β-actin (Cat. No. 664804) was used as a loading control at a 1:25000 dilution. Western-Ready™ ECL Substrate Premium Kit (Cat. No. 426319) was used as a detection agent. Lane M: Molecular weight marker.
  • B. W22030A_PURE_PRAS40_ICC_072224
    HeLa cells were fixed with fixation buffer (Cat. No. 420801) for 10 minutes, permeabilized with ice-cold methanol, and then blocked with 5% FBS for 1 hour at room temperature. Cells were then intracellularly stained with Purified anti-PRAS40/AKT1S1 (clone W22030A), followed by incubation with Alexa Fluor® 594 Goat anti-rat IgG (Cat. No. 405422), for 1 hour at room temperature. Nuclei were counterstained with DAPI (Cat. No. 422801). The image was captured with a 40X objective. Scale bar: 25 µm
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667651 25 µg 132€
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667652 100 µg 332€
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Description

Numerous growth factors and hormones have the capability to initiate the phosphoinositide 3-kinase signaling pathway, leading to the activation of downstream effector proteins such as the serine/threonine kinase AKT. A 40 kDa proline-rich protein called PRAS40 (also known as AKT1S1), which interacts with 14-3-3 proteins, is one known AKT substrate. In order to transduce AKT signals to the mTOR complex, PRAS40/AKT1S1 interacts with mTOR which regulates cell growth and cell cycle and acts as a negative regulator of autophagy. Inhibition of mTOR signaling promotes PRAS40/AKT1S1 binding to mTOR, which reduces mTOR activity. In PC12 cells, treatment with insulin-like growth factor-1 (IGF1) can cause phosphorylation of PRAS40/AKT1S1 by the PI3K/AKT signaling pathway. In insulin-deficient cells, PRAS40/AKT1S1 interacts with raptor in mTOR complex 1 (mTORC1) and prevents the cell cycle protein Rheb from activating the mTORC1 pathway. The inhibition of mTORC1 by PRAS40/AKT1S1 is relieved when AKT phosphorylates PRAS40/AKT1S1 at Thr246. Additionally, PRAS40/AKT1S1 is phosphorylated on Ser183, Ser212, and Ser221 by mTORC1, which releases it from mTORC1.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human, Mouse, Rat
Antibody Type
Monoclonal
Host Species
Rat
Immunogen
Recombinant human PRAS40/AKT1S1 protein
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

WB - Quality tested
ICC - Verified
Recommended Usage

Each lot of this antibody is quality control tested by western blotting. For western blotting, the suggested use of this reagent is 0.125 - 1.0 µg/mL. For immunocytochemistry, a concentration range of 1.0 - 5.0 μg/mL is recommended. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

For use of this antibody (clone W22030A) in immunocytochemistry (ICC), 4% PFA fixation followed by permeabilization with ice-cold methanol is recommended.

This antibody (clone W22030A) is not suitable for use in immunohistochemistry (IHC).

Additional Product Notes

This antibody has been tested in knockout/knockdown models for WB and ICC.

Antigen Details

Structure
PRAS40/AKT1S1 is a 256 amino acid protein with a predicted molecular weight of 27 kD.
Distribution

Widely expressed with highest levels of expression in the liver and heart

Function
Negative regulator of mTORC1 and plays a role in nerve growth factor-mediated neuroprotection
Interaction
mTORC1, MTOR, MLST8, RPTOR
Cell Type
Neurons
Biology Area
Cell Biology, Cell Proliferation and Viability, Neuroscience, Signal Transduction
Molecular Family
Adaptor Proteins
Antigen References
  1. Cantley LC. 2002. Science. 296: 1655-7.
  2. Kovacina KS, et al. 2003. J Biol Chem. 278:10189-94.
  3. Vander Haar E, et al. 2007. Nat Cell Biol. 9:316-23.
  4. Sancak Y, et al. 2007. Mol Cell. 25:903-15.
  5. Oshiro N, et al. 2007. J Biol Chem. 282:20329-39.
  6. Wang L, et al. 2008. J Biol Chem. 283:15619-27.
  7. Wang H, et al. 2012. Neurosci Lett. 516:105-9.
Gene ID
84335 View all products for this Gene ID
UniProt
View information about PRAS40 on UniProt.org
Go To Top Version: 1    Revision Date: 07.22.2024

For Research Use Only. Not for diagnostic or therapeutic use.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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