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> Phospho-mTOR (S2448) ELISA Kit

Phospho-mTOR (S2448) ELISA Kit

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Cat# PEL-mTOR-S2448-1

Size : 1PlateKit

Brand : RayBiotech

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Phone : +1 850 650 7790

Phospho-mTOR (S2448) ELISA KitRayBio® Phospho-mTOR (Ser2448) ELISA Kit was used to measure mTOR activation in gastrocnemius muscle extracts from WT and dyW−/− mice following vemurafenib treatment. Figure adapted from Oliveira-Santos et al., Disease Models & Mechanisms, 2023.RayBio® Phospho-mTOR (Ser2448) and Total mTOR ELISA Kits assessed increased mTOR phosphorylation in irradiated mouse hearts, indicating activation of the mTOR signaling pathway. Figure adapted from Azimzadeh et al., Cancers, 2023.Phospho-mTOR (S2448) ELISA Kit

Phospho-mTOR (S2448) ELISA Kit

Phospho-mTOR (S2448) ELISA Kit

The RayBiotech Phospho-mTOR (S2448) ELISA Kit measures phosphorylated mTOR (Ser2448) and total (pan) mTOR in cell and tissue lysates.
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Product Description

Specifications

Size1 Plate Kit, 2 Plate Kit, 5 Plate Kit
SpeciesHuman, Mouse
Specificity
This ELISA kit recognizes Human
Compatible Sample TypesTissue Lysates, Cell Lysates
Accession Number
P42345|Q9JLN9
Gene Id
2475
Gene Symbols
MTOR, FRAP, FRAP1, FRAP2, RAFT1, RAPT1
Protein Name / Synonyms
Serine/threonine-protein kinase mTOR (EC 2.7.11.1) (FK506-binding protein 12-rapamycin complex-associated protein 1) (FKBP12-rapamycin complex-associated protein) (Mammalian target of rapamycin) (mTOR) (Mechanistic target of rapamycin) (Rapamycin and FKBP12 target 1) (Rapamycin target protein 1)
Quantitative/Semi-QuantitativeSemi-Quantitative
Solid Support
96-well Microplate
Design PrincipleSandwich-based
Method Of DetectionColorimetric
Research AreaPost-Translational Modifications, Phosphorylation, AMPK Signaling, mTOR Signaling, PI3K-AKT Signaling, Autophagy
Shipping TypeBlue ice
Storage-20°C

Product Features

  • Simultaneously measure phosphorylated protein and pan protein in one experiment (for normalization purpose)
  • Screen numerous different cell lysates without performing a Western Blot analysis
  • Minimal hands-on time, convenient, and non-radioactive material

Kit Components

  • Pre-Coated 96-well Strip Microplate
  • Wash Buffer
  • Anti-Phospho Antibody
  • Anti-Pan Antibody (*not included in phospho-only version)
  • HRP-Conjugated Secondary Antibody
  • Assay Diluent
  • TMB One-Step Substrate
  • Stop Solution
  • Lysis Buffer
  • Positive Control Sample

Additional Materials Required

  • 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

Assay Procedure Summary

  1. Prepare all reagents and samples as instructed in the manual.
  2. Add 100 µl of sample or positive control to each well.
  3. Incubate 2.5 h at RT or O/N at 4 °C.
  4. Add 100 µl of prepared primary antibody to each well.
  5. Incubate 1 h at RT.
  6. Add 100 µl of prepared 1X HRP-Streptavidin to each well.
  7. Incubate 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.

Typical Data

A431 cells were treated with recombinant human EGF at 37°C for 20 min. Cells were solubilzed at 4 x 107 cells/ml in Cell Lysate Buffer. Serial dilutions of lysates were analyzed in this ELISA (see Reagent Preparation step 4).

293 cells were untreated or treated with 20% FBS for 20 min. Cell lysates were analyzed using this phosphoELISA. A431 cells were treated with recombinant human EGF at 37°C for 20 min. Cell lysates were analyzed using Western Blot. 

RayBio® Phospho-mTOR (Ser2448) ELISA Kit was used to measure mTOR activation in gastrocnemius muscle extracts from WT and dyWˆ’/ˆ’ mice following vemurafenib treatment. Figure adapted from Oliveira-Santos et al., Disease Models & Mechanisms, 2023.

RayBio® Phospho-mTOR (Ser2448) and Total mTOR ELISA Kits assessed increased mTOR phosphorylation in irradiated mouse hearts, indicating activation of the mTOR signaling pathway. Figure adapted from Azimzadeh et al., Cancers, 2023.

Storage/Stability

Upon receipt, the kit should be stored at €“20°C. Please use within 6 months from the date of shipment.

citations

  • Late Effects of Chronic Low Dose Rate Total Body Irradiation on the Heart Proteome of ApoE−/− Mice Resemble Premature Cardiac Ageing

    Species:
    Mouse
    Sample Type:
    heart
    Year:
    2023
    Journal Name:
    Cancers

  • Vemurafenib improves muscle histopathology in a mouse model of LAMA2-related congenital muscular dystrophy

    Year:
    2023
    Journal Name:
    Disease Models & Mechanisms
    PubMed ID:
    37021539

  • IGF2 role in adrenocortical carcinoma biology

    Species:
    Human
    Sample Type:
    Other / Unspecified (protein extracts of H295R cells)
    Year:
    2019
    Journal Name:
    Endocrine
    PubMed ID:
    31378849

  • Lysophosphatidic acid represses autophagy in prostate carcinoma cells

    Year:
    2019
    Journal Name:
    Biochemistry and Cell Biology-biochimie Et Biologie Cellulaire
    PubMed ID:
    30403494

  • Aegle marmelos differentially affects hepatic markers of glycolysis, insulin signalling pathway, hypoxia, and inflammation in HepG2 cells grown in fructose versus glucose-rich environment.

    Species:
    human
    Sample Type:
    Cell Lysate (human heptaocellular)
    Year:
    2018
    Journal Name:
    Molecular and Cellular Biochemistry
    PubMed ID:
    28766170

  • Neuroprotective Effect of Fisetin Through Suppression of IL-1R/TLR Axis and Apoptosis in Pentylenetetrazole-Induced Kindling in Mice

    Year:
    2021
    Journal Name:
    Frontiers in neurology
    PubMed ID:
    34354662

  • Dietary polyphenols promote resilience against sleep deprivation-induced cognitive impairment by activating protein translation

    Year:
    2018
    Journal Name:
    FASEB J
    PubMed ID:
    29702026

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