MTOR ELISA Kit (Mechanistic Target of Rapamycin (serine/threonine Kinase))

Details for Product FRAP1 ELISA Kit No. ABIN2748394, Supplier: Log in to see
Antigen
  • FRAP1
  • FRAP
  • FRAP2
  • RAFT1
  • RAPT1
  • 2610315D21Rik
  • AI327068
  • Frap1
  • flat
  • frap1
  • tor
  • wu:fc22h08
  • mechanistic target of rapamycin (serine/threonine kinase)
  • MTOR
  • Mtor
  • mtor
Alternatives
Human MTOR ELISA Kit
Epitope
pSer2448
Reactivity
Human
Alternatives
Kits with alternative reactivity to:
23
15
12
1
1
1
1
1
1
1
1
Method Type
Cell ELISA
Application
ELISA
Options
Supplier
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Supplier Product No.
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Purpose Cell-Based Human mTOR (S2448) Phosphorylation ELISA Kit. Suitable for adherent whole cell lines.
Sample Type Cell Culture Cells
Analytical Method Semi-Quantitative
Detection Method Colorimetric
Specificity The antibodies provided in this kit recognizes human and mouse mTOR phosphorylated at Serine-2448 and total mTOR for comparison.
Characteristics
  • Site and signal pathway-specific
  • In vitro detection of adherent cell culture
  • No sample lysis needed
  • Compatible with a standard ELISA plate reader
  • Faster results than with ELISA
  • Adaptable for high-throughput screening and drug discovery
Components
  • uncoated 96-well Microplate
  • Wash Buffer A
  • Wash Buffer B
  • Fixing Solution
  • Quenching Buffer
  • Blocking Buffer
  • Anti-phospho antibody
  • Anti-pan antibody
  • HRP-Conjugated Secondary Antibody
  • TMB One-Step Substrate
  • Stop Solution
  • Material not included
    • Distilled or deionized water
    • 100 mL and 1 liter graduated cylinders
    • Tubes to prepare sample dilutions
    • Protease and Phosphatase inhibitors
    • Precision pipettes to deliver 2 μL to 1 mL volumes
    • Adjustable 1-25 mL pipettes for reagent preparation
    • Benchtop rocker or shaker
    • Microplate reader capable of measuring absorbance at 450 nm
    Alternative Name mTOR (FRAP1 ELISA Kit Abstract)
    Background Mechanistic target of rapamycin, mammalian target of rapamycin (mTOR), FK506-binding protein 12-rapamycin-associated protein 1 (FRAP1)
    Gene ID 2475
    UniProt P42345
    Research Area Cardiovascular, Cardiogenesis, Cell Cycle, DNA/RNA
    Pathways PI3K-Akt Signaling, RTK Signaling, AMPK Signaling, Interferon-gamma Pathway, Fc-epsilon Receptor Signaling Pathway, EGFR Signaling Pathway, Neurotrophin Signaling Pathway, Regulation of Actin Filament Polymerization, Regulation of Muscle Cell Differentiation, Regulation of Cell Size, Skeletal Muscle Fiber Development, Regulation of Carbohydrate Metabolic Process, Autophagy, CXCR4-mediated Signaling Events
    Sample Volume 100 μL
    Plate Uncoated
    Protocol
    1. Seed 10,000-30,000 cells into each well and incubate overnight.
    2. Apply various treatment, inhibitors or activators according to manufacture's instructions.
    3. Add 100 μL of Fixing Solution into each well and incubate for 20 min at RT with shaking.
    4. Add 200 μL of prepared 1X Quenching Buffer and incubate 20 min at RT.
    5. Add 200 μL of Blocking Solution and incubate for 1 h at 37 °C.
    6. Add 50 μL of 1X anti-phospho-protein specific antibody or anti-pan-protein specific antibody to each well and incubate for 2 h at RT.
    7. Add 50 μL of prepared 1X HRP-Anti-Rabbit or Mouse IgG and incubate for 1 h at RT.
    8. Add 100 μL of TMB One-Step Substrate Reagent to each well.
    9. Incubate 30 min at RT.
    10. Add 50 μL of Stop Solution to each well.
    11. Read at 450 nm immediately.
    Assay Procedure

    Seed 10,000-30,000 cells into each well and incubate overnight.
    Apply various treatment, inhibitors or activators according to manufacture's instructions.
    Add 100 μL of Fixing Solution into each well and incubate for 20 min at RT with shaking.
    Add 200 μL of prepared 1X Quenching Buffer and incubate 20 min at RT.
    Add 200 μL of Blocking Solution and incubate for 1 h at 37 °C.
    Add 50 μL of 1X anti-phospho-protein specific antibody or anti-pan-protein specific antibody to each well and incubate for 2 h at RT.
    Add 50 μL of prepared 1X HRP-Anti-Rabbit or Mouse IgG and incubate for 1 h at RT.
    Add 100 μL of TMB One-Step Substrate Reagent to each well.
    Incubate 30 min at RT.
    Add 50 μL of Stop Solution to each well.
    Read at 450 nm immediately.

    Restrictions For Research Use only
    Storage -20 °C
    Storage Comment The entire kit may be stored at -20°C for up to 6 months from the date of shipment. Avoid repeated freeze-thaw cycles.
    Expiry Date 6 months
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