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Product last modified at: 2024-11-25T23:15:09.113Z
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PDP - Template Name: Monoclonal Antibody (Alexa Fluor Conjugate)
PDP - Template ID: *******c8ce56b
  1. Products /
  2. Antibody Conjugates /
  3. Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate)
R Recombinant
Recombinant: Superior lot-to-lot consistency, continuous supply, and animal-free manufacturing.

Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate) #4075

Filter:
  • F

    Supporting Data

    REACTIVITY H M R Hm Mk Dm Z B
    SENSITIVITY Endogenous
    MW (kDa)
    Source/Isotype Rabbit IgG
    Application Key:
    • F-Flow Cytometry 
    Species Cross-Reactivity Key:
    • H-Human 
    • M-Mouse 
    • R-Rat 
    • Hm-Hamster 
    • Mk-Monkey 
    • Dm-D. melanogaster 
    • Z-Zebrafish 
    • B-Bovine 

    Product Information

    Product Description

    This Cell Signaling Technology antibody is conjugated to Alexa Fluor® 647 fluorescent dye and tested in-house for direct flow cytometric analysis of human cells. The unconjugated antibody #4060 reacts with human, mouse, rat, hamster, bovine, D. melanogaster and zebra fish Phospho-Akt protein. CST expects that Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate) will also recognize Phospho-Akt in these species.

    Product Usage Information

    Application Dilution
    Flow Cytometry (Fixed/Permeabilized) 1:50

    Storage

    Supplied in PBS (pH 7.2), less than 0.1% sodium azide and 2 mg/ml BSA. Store at 4°C. Do not aliquot the antibody. Protect from light. Do not freeze.

    Protocol

    Specificity / Sensitivity

    Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate) detects endogenous levels of Akt only when phosphorylated at Ser473.

    Species Reactivity:

    Human, Mouse, Rat, Hamster, Monkey, D. melanogaster, Zebrafish, Bovine

    The antigen sequence used to produce this antibody shares 100% sequence homology with the species listed here, but reactivity has not been tested or confirmed to work by CST. Use of this product with these species is not covered under our Product Performance Guarantee.

    Species predicted to react based on 100% sequence homology:

    Chicken, Xenopus, Dog, Pig

    Source / Purification

    Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues around Ser473 of human Akt. The antibody was conjugated to Alexa Fluor® 647 under optimal conditions with an F/P ratio of 2-6. The Alexa Fluor® 647 dye is maximally excited by red light (e.g. 633 nm He-Ne laser). Antibody conjugates of the Alexa Fluor® 647 dye produce bright far-red-fluorescence emission, with a peak at 665 nm.

    Background

    Akt, also referred to as PKB or Rac, plays a critical role in controlling cell survival and apoptosis (1-3). This protein kinase is activated by insulin and various growth and survival factors to function in a wortmannin-sensitive pathway involving PI3 kinase (2,3). Akt is activated by phospholipid binding and activation loop phosphorylation at Thr308 by PDK1 (4) and by phosphorylation within the carboxy terminus at Ser473. The previously elusive PDK2 responsible for phosphorylation of Akt at Ser473 has been identified as mammalian target of rapamycin (mTOR) in a rapamycin-insensitive complex with rictor and Sin1 (5,6). Akt promotes cell survival by inhibiting apoptosis through phosphorylation and inactivation of several targets, including Bad (7), forkhead transcription factors (8), c-Raf (9), and caspase-9. PTEN phosphatase is a major negative regulator of the PI3K/Akt signaling pathway (10). LY294002 is a specific PI3 kinase inhibitor (11). Another essential Akt function is the regulation of glycogen synthesis through phosphorylation and inactivation of GSK-3α and β (12,13). Akt may also play a role in insulin stimulation of glucose transport (12). In addition to its role in survival and glycogen synthesis, Akt is involved in cell cycle regulation by preventing GSK-3β-mediated phosphorylation and degradation of cyclin D1 (14) and by negatively regulating the cyclin-dependent kinase inhibitors p27 Kip1 (15) and p21 Waf1/Cip1 (16). Akt also plays a critical role in cell growth by directly phosphorylating mTOR in a rapamycin-sensitive complex containing raptor (17). More importantly, Akt phosphorylates and inactivates tuberin (TSC2), an inhibitor of mTOR within the mTOR-raptor complex (18,19).
    1. Franke, T.F. et al. (1997) Cell 88, 435-7.
    2. Burgering, B.M. and Coffer, P.J. (1995) Nature 376, 599-602.
    3. Franke, T.F. et al. (1995) Cell 81, 727-36.
    4. Alessi, D.R. et al. (1996) EMBO J 15, 6541-51.
    5. Sarbassov, D.D. et al. (2005) Science 307, 1098-101.
    6. Jacinto, E. et al. (2006) Cell 127, 125-37.
    7. Cardone, M.H. et al. (1998) Science 282, 1318-21.
    8. Brunet, A. et al. (1999) Cell 96, 857-68.
    9. Zimmermann, S. and Moelling, K. (1999) Science 286, 1741-4.
    10. Cantley, L.C. and Neel, B.G. (1999) Proc Natl Acad Sci USA 96, 4240-5.
    11. Vlahos, C.J. et al. (1994) J Biol Chem 269, 5241-8.
    12. Hajduch, E. et al. (2001) FEBS Lett 492, 199-203.
    13. Cross, D.A. et al. (1995) Nature 378, 785-9.
    14. Diehl, J.A. et al. (1998) Genes Dev 12, 3499-511.
    15. Gesbert, F. et al. (2000) J Biol Chem 275, 39223-30.
    16. Zhou, B.P. et al. (2001) Nat Cell Biol 3, 245-52.
    17. Navé, B.T. et al. (1999) Biochem J 344 Pt 2, 427-31.
    18. Inoki, K. et al. (2002) Nat Cell Biol 4, 648-57.
    19. Manning, B.D. et al. (2002) Mol Cell 10, 151-62.

    Database Links

    UniProt ID: P31751 , Q9Y243 , P31749


    Entrez-Gene Id: 208 , 10000 , 207


    Limited Uses

    Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

    Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

    For Research Use Only. Not For Use In Diagnostic Procedures.
    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    XP is a registered trademark of Cell Signaling Technology, Inc.
    This product is provided under an intellectual property license from Life Technologies Corporation. The transfer of this product is conditioned on the buyer using the purchased product solely in research conducted by the buyer, excluding contract research or any fee for service research, and the buyer must not (1) use this product or its components for (a) diagnostic, therapeutic or prophylactic purposes; (b) testing, analysis or screening services, or information in return for compensation on a per-test basis; or (c) manufacturing or quality assurance or quality control, and/or (2) sell or transfer this product or its components for resale, whether or not resold for use in research. For information on purchasing a license to this product for purposes other than as described above, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, CA 92008 USA or [email protected].
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