Human/Mouse/Rat Akt Pan Specific Antibody MAB2055: R&D Systems

Human/Mouse/Rat Akt Pan Specific Antibody

(4 citations)
(1 Review)
  
  • Species Reactivity
    Human, Mouse, Rat
  • Specificity
    Detects human, mouse and rat Akt in direct ELISAs and Western blots.
  • Source
    Monoclonal Mouse IgG2B Clone # 281046
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    E. coli-derived recombinant human Akt1
    Ser2-Ala480
    Accession # P31749
  • 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
    0.2 µg/mL
    See below
  • Simple Western
    2 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.
  • Immunocytochemistry
    8-25 µg/mL
    See below
  • 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 Human/Mouse Akt by Western Blot. Western blot shows lysates of MCF-7 human breast cancer cell line, MBA-MB-123 human breast cancer cell line, and TS1 mouse helper T cell line. PVDF membrane was probed with 0.2 µg/mL Mouse Anti-Human/Mouse/Rat Akt Pan Specific Monoclonal Antibody (Catalog # MAB2055) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 4.
Detection of Human Akt Pan Specific by Western Blot. Western blot shows Recombinant Human Active Akt1 (Catalog # 1775-KS), recombinant human Akt2, and recombinant human Akt3 (5 ng/lane). PVDF membrane was probed with 0.2 µg/mL Mouse Anti-Human/Mouse/Rat Akt Pan Specific Monoclonal Antibody (Catalog # MAB2055) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 4.
Immunocytochemistry
Akt in MDA‑MB‑231 Human Cell Line. Akt was detected in immersion fixed MDA‑MB‑231 human breast cancer cell line using Mouse Anti-Human/
Mouse/Rat Akt Monoclonal Antibody (Catalog # MAB2055) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (yellow; Catalog # NL007) and counter­stained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Intracellular Staining by Flow Cytometry
Detection of Akt in MCF‑7 Human Cell Line by Flow Cytometry. MCF‑7 human breast cancer cell line was stained with Mouse Anti-Human/Mouse/Rat Akt Monoclonal Antibody (Catalog # MAB2055, filled histogram) or isotype control antibody (Catalog # MAB0041, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG F(ab')2 Secondary Antibody (Catalog # F0102B). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.
Immunocytochemistry
Akt (pan) in 293T Human Cell Line. Akt pan specific (panels A and C) and Akt1 (E17K Mutation) (panels B and D) were detected in immersion fixed 293T human embryonic kidney cell line transfected with wild type (panels A and B) or E17K mutated (panels C and D) Akt1 using Mouse Anti-Human/Mouse/Rat Akt Pan Specific Monoclonal Antibody (Catalog # MAB2055) and Mouse Anti-Human Akt1 (E17K Mutation) Monoclonal Antibody (Catalog # MAB6815). Both antibodies were used at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to plasma membranes and cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Detection of Human Akt by Simple WesternTM. Simple Western lane view shows lysates of MCF‑7 human breast cancer cell line, loaded at 0.2 mg/mL. A specific band was detected for Akt at approximately 62 kDa (as indicated) using 2 µg/mL of Mouse Anti-Human/Mouse/Rat Akt Pan Specific Monoclonal Antibody (Catalog # MAB2055). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.5 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

Akt, also known as protein kinase B (PKB), is a central kinase in such diverse cellular processes as glucose uptake, cell cycle progression, and apoptosis. Three highly homologous members define the Akt family: Akt1 (PKB alpha ), Akt2 (PKB beta ), and Akt3 (PKB gamma ). All three Akts contain an amino-terminal pleckstrin homology domain, a central kinase domain, and a carboxyl-terminal regulatory domain.

  • 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.

4 Citations: Showing 1 - 4
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Species
Applications
Sample Type
  1. Lack of CD2AP disrupts Glut4 trafficking and attenuates glucose uptake in podocytes.
    Authors: Tolvanen T, Dash S, Polianskyte-Prause Z, Dumont V, Lehtonen S
    J Cell Sci, 2015;128(24):4588-600.
    Species: Mouse
    Sample Type: Cell Lysates
    Application: WB
  2. 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
  3. 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
  4. 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
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Rat Anti-Mouse IgG2B APC-conjugated Antibody

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Mouse IgG VisUCyte HRP Polymer Antibody

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Mouse IgG2B Antibody

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Average Rating: 4 (Based on 1 review)

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We have 1 review tested in 1 species: Human.
We have 1 review tested in 1 application: Immunocytochemistry/Immunofluorescence.

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 ICC/IF Human Anonymous 11/03/2017
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Summary

ApplicationImmunocytochemistry/Immunofluorescence
Sample TestedProstate tissue
SpeciesHuman

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