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Purified anti-AKT Phospho (Ser473) Antibody

Pricing & Availability
Clone
Poly23A04 (See other available formats)
Regulatory Status
RUO
Other Names
Protein kinase B alpha, PRK-BA, PKB, Serine/Threonine specific kinase RAC alpha, Protein kinase Akt, RAC
Isotype
Rabbit Polyclonal IgG
Ave. Rating
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Product Citations
publications
Poly23A04_PURE_AKT_WB_010424
Whole cell extracts (15 µg total protein) from serum-starved HeLa cells untreated (-) or treated (+) with IGF-1 (100 ng/mL, 30 min), and from serum-starved NIH/3T3 cells untreated (-) or treated (+) with PDGF-BB (200 ng/mL, 15 min) were resolved on a 4-12% Bis-Tris gel, transferred to a PVDF membrane, and probed with 1 µg/mL (1:1000 dilution) of Purified anti-AKT Phospho (Ser473) (clone Poly23A04) for 2 hours at room temperature (1st and 3rd blots). Proteins were visualized by chemiluminescence detection using HRP Goat anti-rabbit IgG (Cat. No. 406401) at a 1:3000 dilution. Equal AKT Phospho (Ser473) loading was confirmed by probing membranes with pan Purified anti-AKT1 (clone O94E10) (Cat. No. 680302) at a 1:500 dilution (2nd and 4th blots). Direct-Blot™ HRP anti-GAPDH (Cat. No. 607904) was used as a loading control at a 1:50000 dilution. Western-Ready™ ECL Substrate Plus Kit (Cat. No. 426317) was used as a detection agent. Lane M: Molecular weight marker
  • Poly23A04_PURE_AKT_WB_010424
    Whole cell extracts (15 µg total protein) from serum-starved HeLa cells untreated (-) or treated (+) with IGF-1 (100 ng/mL, 30 min), and from serum-starved NIH/3T3 cells untreated (-) or treated (+) with PDGF-BB (200 ng/mL, 15 min) were resolved on a 4-12% Bis-Tris gel, transferred to a PVDF membrane, and probed with 1 µg/mL (1:1000 dilution) of Purified anti-AKT Phospho (Ser473) (clone Poly23A04) for 2 hours at room temperature (1st and 3rd blots). Proteins were visualized by chemiluminescence detection using HRP Goat anti-rabbit IgG (Cat. No. 406401) at a 1:3000 dilution. Equal AKT Phospho (Ser473) loading was confirmed by probing membranes with pan Purified anti-AKT1 (clone O94E10) (Cat. No. 680302) at a 1:500 dilution (2nd and 4th blots). Direct-Blot™ HRP anti-GAPDH (Cat. No. 607904) was used as a loading control at a 1:50000 dilution. Western-Ready™ ECL Substrate Plus Kit (Cat. No. 426317) was used as a detection agent. Lane M: Molecular weight marker
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627451 50 µL 168€
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627452 200 µL 420€
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Description

AKT (also known as protein kinase B alpha) is a 60 kD serine/threonine-specific kinase containing a pleckstrin domain. AKT plays a critical role in controlling survival and apoptosis. This kinase is ubiquitously expressed and translocates to the membrane upon cell activation. It is activated by insulin and various growth and survival factors to function in a wortmannin-sensitive pathway involving PI3-kinase. AKT is activated by phospholipid binding and activation loop phosphorylation at Thr308 by PDK1 and by phosphorylation within the carboxy terminus at Ser473. The previously elusive PDK2, responsible for phosphorylation of AKT at Ser473, has been identified as a mammalian target of rapamycin (mTOR) in a rapamycin-insensitive complex with RICTOR and SIN1. Activated AKT phosphorylates a wide range of substrates including transcription factors (e.g. FOXO1), kinases (GSK-3beta, Raf-1, ASK, Chk1) and other proteins with important signaling roles (e.g. Bad, MDM2). AKT promotes cell survival by inhibiting apoptosis through phosphorylation of several targets, including Bad, forkhead transcription factor, c-Raf and caspase-9. AKT also plays a critical role in cell growth by directly phosphorylating mTOR in a rapamycin-sensitive complex containing raptor. More importantly, AKT phosphorylates and inactivates tuberin (TSC2), an inhibitor of mTOR within the mTOR-raptor complex.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human, Mouse
Antibody Type
Polyclonal
Host Species
Rabbit
Immunogen
Synthetic peptide of human AKT phosphorylated at Ser473
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
Preparation
The antibody was purified by affinity chromatography.
Concentration
Lot-specific (to obtain lot-specific concentration and expiration, please enter the lot number in our Certificate of Analysis online tool.)
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

WB - Quality tested
Recommended Usage

Each lot of this antibody is quality control tested by western blotting. For western blotting, the suggested dilution of this reagent is 1:1000 - 1:4000. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

This clone was tested by Western Blot using lysates prepared from HeLa cells untreated or treated with IGF-1, and NIH/3T3 cells untreated or treated with PDGF-BB.

RRID
AB_3097484 (BioLegend Cat. No. 627451)
AB_3097484 (BioLegend Cat. No. 627452)

Antigen Details

Structure
AKT is a 541 amino acid protein with a predicted molecular weight of 62 kD
Distribution

Ubiquitously expressed, translocates to the membrane upon activation

Function
Catalytically inactive multimeric complex. AKT is activated by phospholipid binding and activation loop phosphorylation at Thr308 by PDK1 and by phosphorylation within the carboxy terminus at Ser473 by PDK2.
Interaction
Interacts through pleckstrin homology domain with second messengers. Interacts with AKTIP and CDKN1B.
Biology Area
Cell Adhesion, Cell Biology, Cell Death
Molecular Family
Phospho-Proteins, Protein Kinases/Phosphatase
Antigen References
  1. Jacinto E, et al. 2006. Cell. 127:125-37.
  2. Inoki K, et al. 2002. Nat Cell Biol. 4:648-57.
  3. Franke TF, et al. 1995. Cell. 81:727-36.
  4. Brunet A, et al. 1999. Cell. 96:857-68.
  5. Yang WL, et al. 2010. Cell Cycle. 9:487-97.
Gene ID
207 View all products for this Gene ID
UniProt
View information about AKT on UniProt.org
Go To Top Version: 1    Revision Date: 01-04-2024

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

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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Toll-Free Phone: 1-877-Bio-Legend (246-5343) Phone: (858) 768-5800 Fax: (877) 455-9587

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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