Human/Mouse Phospho-PRAS40 (T246) Antibody

(2 citations)   
  • Species Reactivity
    Human, Mouse
  • Specificity
    Detects human and mouse PRAS40 when phosphorylated at T246
  • Source
    Monoclonal Mouse IgG2A Clone # 760502
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    Phosphopeptide containing the human PRAS40 T246 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
  • Immunocytochemistry
    8-25 µg/mL
    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 and Mouse Phospho-PRAS40 (T246) by Western Blot. Western blot shows lysates of MCF‑7 human breast cancer cell line and NIH‑3T3 mouse embryonic fibroblast cell line untreated (-) or treated (+) with 100 ng/mL Recombinant Human IGF‑I (Catalog # 291-G1) for 20 minutes and 10 ng/mL Recombinant Human PDGF‑BB (Catalog # 220-BB) for 5 minutes. PVDF membrane was probed with 0.5 µg/mL of Mouse Anti-Human Phospho-PRAS40 (T246) Monoclonal Antibody (Catalog # MAB6890) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for Phospho-PRAS40 (T246) at approximately 40 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Immunocytochemistry
Phospho-PRAS40 (T246) in MCF‑7 Human Cell Line. PRAS40 phosphorylated at T246 was detected in immersion fixed MCF‑7 human breast cancer cell line using Mouse Anti-Human/Mouse Phospho-PRAS40 (T246) Monoclonal Antibody (Catalog # MAB6890) at 25 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 cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Preparation and Storage
  • Reconstitution
    Sterile PBS to a final concentration of 0.5 mg/mL.
  • 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: PRAS40

PRAS40 (Proline-rich AKT1 substrate 1; also Akt1S1 and p39) is a 40-42 kDa cytoplasmic phosphoprotein that lacks generally recognized structural motifs. It is widely expressed and is considered to be key regulator of mTORC1 (mTOR plus raptor and G beta L), a complex through which Akt signals into the cell. Through phosphorylation, mTORC1 activity is upregulated by PRAS40. In particular, nonphosphorylated PRAS40 binds to and serves as a negative regulator of mTORC1 activity. Upon Insulin signaling, PRAS40 is phosphorylated on Thr246, Ser221 and Ser183. This causes it to bind 14-3-3 and results in its dissociation from mTORC1, freeing up mTOR to regulate (positively or negatively) protein synthesis. Human PRAS40 is 256 amino acids (aa) in length. It contains one extended Pro-rich region (aa 35-96) plus at least nine utilized Ser/Thr phosphorylation sites. There is one alternative start site at Met131. Over aa 119-256, human PRAS40 shares 93% aa identity with mouse PRAS40.

  • Long Name:
    40 kDa Proline-rich Akt1 Substrate
  • Entrez Gene IDs:
    84335 (Human); 67605 (Mouse); 292887 (Rat)
  • Alternate Names:
    40 kDa; AKT1 substrate 1 (proline-rich); AKT1S1; Lobe; MGC2865; PRAS40; proline-rich AKT1 substrate 1,40 kDa proline-rich AKT substrate
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.

2 Citations: Showing 1 - 2
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Species
Applications
Sample Type
  1. Soluble bone-derived osteopontin promotes migration and stem-like behavior of breast cancer cells
    Authors: GM Pio, Y Xia, MM Piaseczny, JE Chu, AL Allan
    PLoS ONE, 2017;12(5):e0177640.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  2. Targeted depletion of PIK3R2 induces regression of lung squamous cell carcinoma
    Oncotarget, 2016;0(0):.
    Species: Human
    Sample Type: Cell Culture Supernates
    Application: WB
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Mouse IgG2A Isotype Control

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Secondary Antibodies
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Mouse IgG HRP-conjugated Antibody

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Donkey Anti-Mouse IgG NorthernLights™ NL637-conjugated Antibody

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Mouse F(ab)2 IgG (H+L) APC-conjugated Antibody

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Donkey Anti-Mouse IgG NorthernLights™ NL493-conjugated Antibody

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Mouse F(ab)2 IgG (H+L) Fluorescein-conjugated Antibody

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Mouse IgG Antibody

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Goat Anti-Mouse IgG Biotinylated Antibody

WB BAF007 4
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Mouse/Rabbit IgG VisUCyte HRP Polymer Antibody

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Rat Anti-Mouse IgG2A PE-conjugated Antibody

Flow F0129 1  
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Mouse IgG VisUCyte HRP Polymer Antibody

IHC VC001  
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Goat Anti-Mouse IgG Fc Affinity Purified Polyclonal Ab

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Mouse F(ab)2 IgG (H+L) PerCP-conjugated Antibody

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Rat Anti-Mouse IgG2A APC-conjugated Antibody

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Mouse IgG2A Antibody

Flow MAB0032  
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Rat Anti-Mouse IgG2A PerCP-conjugated Antibody

Flow F0131  
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