Human/Mouse/Rat Phospho-Akt (S473) Pan Specific Antibody

(16 citations)   
  • Species Reactivity
    Human, Mouse, Rat
  • Specificity
    Detects human, mouse and rat Akt1, Akt2 and Akt3, when phosphorylated at S473, S474 and S472, respectively.
  • Source
    Polyclonal Rabbit IgG
  • Purification
    Antigen and protein A Affinity-purified
  • Immunogen
    Phosphopeptide containing human, mouse, and rat Akt1 S473 site
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.5 µg/mL
    See below
  • Simple Western
    5 µg/mL
    See below
  • Immunohistochemistry
    5-15 µg/mL
    See below
  • CyTOF-ready
    Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
  • Intracellular Staining by Flow Cytometry
    2.5 µg/106 cells
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples

Detection of Mouse and Human Phospho-Akt (S473) Pan Specific by Western Blot. Western blot shows lysates of NIH‑3T3 mouse embryonic fibroblast cell line untreated (-) or treated (+) with 100 ng/mL Human PDGF (Catalog # 120‑HD) for 15 minutes and MCF‑7 human breast cancer cell line untreated or treated with 100 ng/mL Recombinant Human IGF-1 (Catalog # 291‑G1) for 15 minutes. PVDF membrane was probed with 0.5 µg/mL of Rabbit Anti-Human/Mouse/Rat Phospho-Akt (S473) Pan Specific Antigen Affinity-purified Polyclonal Antibody (Catalog # AF887), followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Phospho-Akt (S473) Pan Specific at approximately 60 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Immunohistochemistry

Akt in Human Breast Cancer Tissue. Akt phosphorylated at S473 was detected in immersion fixed paraffin-embedded sections of human breast cancer tissue using Rabbit Anti-Human/Mouse/Rat Phospho-Akt (S473) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF887) at 10 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Rabbit HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS005) and counterstained with hematoxylin (blue). Lower panel shows a lack of labeling if primary antibodies are omitted and tissue is stained only with secondary antibody followed by incubation with detection reagents. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Intracellular Staining by Flow Cytometry

Detection of Phospho-Akt (S473) in IGF-1-treated MCF‑7 cells by Flow Cytometry. Serum-starved MCF‑7 human breast cancer cells were unstimulated (light orange filled histogram) or activated with 100 ng/mL Recombinant human IGF-1 (Catalog # 291-G1, dark orange filled histogram) for 15 minutes, then stained with Rabbit Anti-Human/Mouse/Rat Phospho‑Akt (S473) Antigen Affinity‑purified Polyclonal Antibody (Catalog # AF887) or control antibody (Catalog # AB-105-C, open histogram), followed by Allophycocyanin-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # F0111). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.

Detection of Human Phospho-Akt (S473) by Simple WesternTM. Simple Western lane view shows lysates of MCF‑7 human breast cancer cell line and A549 human lung carcinoma cell line untreated (-) or treated (+) with 100 ng/mL Recombinant Human IGF‑I (Catalog # 291-G1) for 15 minutes, loaded at 0.2 mg/mL. A specific band was detected for Phospho-Akt (S473) at approximately 60 kDa (as indicated) using 5 µg/mL of Rabbit Anti-Human/Mouse/Rat Phospho-Akt (S473) Pan Specific Antigen Affinity-purified Polyclonal Antibody (Catalog # AF887). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 12 months from date of receipt, -20 to -70 °C as supplied.
    • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
    • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: Akt
Akt1, Akt2, and Akt3 make up a family of serine/threonine kinases, often activated downstream of growth factor receptors and PI 3-Kinase. Each Akt family member contains an N-terminal pleckstrin homology (PH) domain, a central kinase domain, and a C-terminal regulatory domain. They have putative roles in cell proliferation and survival, and Akt perturbations are thought to play an important role in tumorigenesis.
  • Long Name:
    v-Akt Murine Thymoma Viral Oncogene Homolog
  • Alternate Names:
    Akt; PKB; RAC
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

16 Citations: Showing 1 - 10
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Species
Applications
Sample Type
  1. Different dynamics of NLRP3 inflammasome-mediated IL?1? production in GM-CSF- and M-CSF-differentiated human macrophages
    Authors: MM Budai, J T?zsér, S Benk?
    J. Leukoc. Biol, 2017;0(0):.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  2. Ocular Delivery of PACAP1-27 Protects the Retina From Ischemic Damage in Rodents
    Invest. Ophthalmol. Vis. Sci, 2016;57(15):6683-6691.
    Species: Mouse
    Sample Type: Tissue Homogenates
    Application: WB
  3. Dual inhibition of CDK4/Rb and PI3K/AKT/mTOR pathways by ON123300 induces synthetic lethality in mantle cell lymphomas.
    Authors: Divakar S, Ramana Reddy M, Cosenza S, Baker S, Perumal D, Antonelli A, Brody J, Akula B, Parekh S, Premkumar Reddy E
    Leukemia, 2016;30(1):86-93.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  4. Interferon-gamma suppresses intestinal epithelial aquaporin-1 expression via Janus kinase and STAT3 activation.
    Authors: Dicay M, Hirota C, Ronaghan N, Peplowski M, Zaheer R, Carati C, MacNaughton W
    PLoS ONE, 2015;10(3):e0118713.
    Species: Mouse
    Sample Type: Cell Lysates
    Application: WB
  5. Neuronal pentraxin 2 supports clear cell renal cell carcinoma by activating the AMPA-selective glutamate receptor-4.
    Authors: von Roemeling C, Radisky D, Marlow L, Cooper S, Grebe S, Anastasiadis P, Tun H, Copland J
    Cancer Res, 2014;74(17):4796-810.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  6. Water-filtered infrared a irradiation in combination with visible light inhibits acute chlamydial infection.
    Authors: Marti H, Koschwanez M, Pesch T, Blenn C, Borel N
    PLoS ONE, 2014;9(7):e102239.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  7. The role of nuclear beta-catenin accumulation in the Twist2-induced ovarian cancer EMT.
    Authors: Mao Y, Xu J, Li Z, Zhang N, Yin H, Liu Z
    PLoS ONE, 2013;8(11):e78200.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  8. An elaborate regulation of Mammalian target of rapamycin activity is required for somatic cell reprogramming induced by defined transcription factors.
    Authors: He, Jing, Kang, Lan, Wu, Tong, Zhang, Jiqin, Wang, Haitao, Gao, Haibo, Zhang, Yu, Huang, Bo, Liu, Wenqiang, Kou, Zhaohui, Zhang, Hongbing, Gao, Shaorong
    Stem Cells Dev, 2012;21(14):2630-41.
    Species: Mouse
    Sample Type: Cell Lysates
    Application: WB
  9. Single amino Acid substitutions in the chemotactic sequence of urokinase receptor modulate cell migration and invasion.
    Authors: Bifulco, Katia, Longanesi-Cattani, Immacola, Franco, Paola, Pavone, Vincenzo, Mugione, Pietro, Di Carluccio, Gioconda, Masucci, Maria Te, Arra, Claudio, Pirozzi, Giuseppe, Stoppelli, Maria Pa, Carriero, Maria Vi
    PLoS ONE, 2012;7(9):e44806.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  10. EphB receptors trigger Akt activation and suppress Fas receptor-induced apoptosis in malignant T lymphocytes.
    Authors: Maddigan A, Truitt L, Arsenault R
    J. Immunol., 2011;187(11):5983-94.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  11. Overexpressing cellular repressor of E1A-stimulated genes protects mesenchymal stem cells against hypoxia- and serum deprivation-induced apoptosis by activation of PI3K/Akt.
    Authors: Deng J, Han Y, Yan C, Tian X, Tao J, Kang J, Li S
    Apoptosis, 2010;15(4):463-73.
    Species: Rat
    Sample Type: Cell Lysates
    Application: WB
  12. Cellular repressor of E1A-stimulated genes inhibits human vascular smooth muscle cell apoptosis via blocking P38/JNK MAP kinase activation.
    Authors: Han Y, Wu G, Deng J, Tao J, Guo L, Tian X, Kang J, Zhang X, Yan C
    J. Mol. Cell. Cardiol., 2010;48(6):1225-35.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  13. The involvement of the fractalkine receptor in the transmigration of neuroblastoma cells through bone-marrow endothelial cells.
    Authors: Nevo I, Sagi-Assif O, Meshel T, Ben-Baruch A, Johrer K, Greil R, Trejo LE, Kharenko O, Feinmesser M, Yron I, Witz IP
    Cancer Lett., 2009;273(1):127-39.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  14. CREG inhibits migration of human vascular smooth muscle cells by mediating IGF-II endocytosis.
    Authors: Han Y, Cui J, Tao J, Guo L, Guo P, Sun M, Kang J, Zhang X, Yan C, Li S
    Exp. Cell Res., 2009;315(19):3301-11.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  15. Osteoblast-derived factors induce an expression signature that identifies prostate cancer metastasis and hormonal progression.
    Authors: Wang G, Haile S, Comuzzi B, Tien AH, Wang J, Yong TM, Jelescu-Bodos AE, Blaszczyk N, Vessella RL, Masri BA, Sadar MD
    Cancer Res., 2009;69(8):3433-42.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  16. Enhancing mammalian target of rapamycin (mTOR)-targeted cancer therapy by preventing mTOR/raptor inhibition-initiated, mTOR/rictor-independent Akt activation.
    Authors: Wang X, Yue P, Kim YA, Fu H, Khuri FR, Sun SY
    Cancer Res., 2008;68(18):7409-18.
    Species: Mouse
    Sample Type: Whole Tissue
    Application: IHC Paraffin-embedded
Expand to show all 16 Citations
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