• 檢測方法Colorimetric
• 精確度

  批次內

  樣品	n	Mean	SD	CV%
  overall	24	?	?	= 4.3%

  批次間

  樣品	n	Mean	SD	CV%
  overall	3	?	?	= 1.8%

• 樣品類型
  Cell Lysate, Tissue Lysate
• 檢測類型Sandwich (quantitative)
• 靈敏度
  = 8 μg/ml
• 范圍
  15.6 μg/ml - 1000 μg/ml
• 實驗步驟Multiple steps standard assay
• 種屬反應性
  與反應:?Mouse, Rat, Human
• 產品概述

  Abcam’s mTOR pSer2448 in vitro ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the accurate quantitative measurement of Ser2448 of mTOR protein in cell and tissue lysates.

  The assay employs an antibody specific to mTOR protein coated onto well plate strips. Samples are pipetted into the wells and mTOR present in the sample is bound to the wells by the immobilized antibody. The wells are washed and an anti-mTOR phospho Ser2448 detector antibody is added. After washing away unbound detector antibody, HRP-conjugated label specific for the detector antibody is pipetted into the wells. The wells are again washed, a TMB substrate solution is added to the wells and blue color develops in proportion to the amount of phosphorylated Ser2448 bound mTOR. The developing blue color is measured at 600 nm. Optionally the reaction can be stopped after 25 – 30 minutes of incubation by adding hydrochloric acid which changes the color from blue to yellow and the intensity can be measured at 450 nm.

• 說明

  Mammalian target of rapamycin (mTOR) is a serine/threonine protein kinase part of two distinct signaling complexes, mTORC1 and mTORC2. These two complexes share four proteins (mTOR, mLST8, DEPTOR, Tti1/tel2), with only mTORC1 containing Raptor and PRAS40 and mTORC2 containing Rictor, mSin1 and Protor1/2. The complex mTORC1 (rapamycin sensitive complex) coordinates inputs from growth factors, stress, energy status, oxygen and amino acids levels to control processes such as protein and lipid synthesis and autophagy. The complex mTORC2 is insensitive to nutrients and rapamycin, but it responds to insulin signaling. It also controls ion transport and cell shape by targeting serum/glucocorticoid protein kinase (SGK1) and protein kinase (PKC-α) respectively.

  The canonical regulation of mTORC1 occurs through the TSC/Rheb pathway which receives signals from AKT, AMPK and IKKβ to activate the complex. Phosphorylation of mTOR at Ser2448 is carried out directly by AKT kinase as well as p70S6 kinase acting as a feedback signal. Phosphorylation at this site is a biomarker for the activation state of the PI-3 kinase pathway as well as the activation status of mTOR. Activation of mTOR leads to phosphorylation of PRAS40, raptor and DEPTOR and the consequential activation of mTORC1. Deregulated signaling of mTOR has been implicated in diseases such as cancer, metabolic syndrome, neurodegeneration and aging. Constitutive activation of PI3K-mTORC1 signaling in cancer cells inhibits autophagy, deregulates protein synthesis via 4E-BP1/eIF4E and increases de novo lipid synthesis via SREBP1. Similarly mTOR signaling is a key factor in the regulation of tissue metabolism in the normal and nutrient overload state affecting the hypothalamus, adipose tissue, the liver, skeletal muscle and pancreas.

• 經測試應用ELISAmore details
• 平臺Microplate

性能

• 存放說明Store at +4°C. Please refer to protocols.

• 組件	1 x 96 tests
  10X Blocking Buffer	1 x 6ml
  10X HRP Label	1 x 1ml
  10X mTOR (pSer2448) Detector Antibody	1 x 700μl
  10X Wash Buffer	1 x 40ml
  Extraction Buffer	1 x 15ml
  mTOR Microplate	1 unit
  Stop Solution	1 x 12ml
  TMB Development Solution	1 x 12ml

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