Phospho-mTOR (S2448) ELISA Kit (ab279868)
Key features and details
- Sample type: Cell Lysate
- Detection method: Colorimetric
- Assay type: Semi-quantitative
- Reacts with: Human
Overview
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Product name
Phospho-mTOR (S2448) ELISA Kit
See all mTOR kits -
Detection method
Colorimetric -
Sample type
Cell Lysate -
Assay type
Semi-quantitative -
Assay duration
Multiple steps standard assay -
Species reactivity
Reacts with: Human -
Product overview
Phospho-mTOR (S2448) ELISA Kit (ab279868) is a very rapid, convenient, and sensitive assay kit that can monitor the activation or function of important biological pathways in human cell lysates. By determining phosphorylated mTOR protein in your experimental model system, you can verify pathway activation in your cell lysates. You can simultaneously measure numerous different cell lysates without spending excess time and effort in performing a Western Blotting analysis.
This Sandwich ELISA kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human phospho-mTOR. An anti-pan mTOR antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and mTOR present in a sample is bound to the wells by the immobilized antibody. The wells are washed and rabbit anti-phospho-mTOR (S2448) antibody is used to detect phosphorylated mTOR. After washing away unbound antibody, HRP-conjugated anti-rabbit IgG is pipetted into the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of mTOR (S2448) bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.
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Notes
Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses. -
Platform
Pre-coated microplate (12 x 8 well strips)
Properties
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Storage instructions
Store at -20°C. Please refer to protocols. -
Components 1 x 96 tests 20X Wash Buffer 1 x 25ml 2X Cell lysate buffer 1 x 10ml 5X Assay Diluent 1 x 15ml HRP-conjugated anti-rabbit IgG concentrate (1000X) 1 vial Pan-mTOR Coated Microplate 1 unit Positive Control - A431 cell lysate 1 vial Rabbit anti-phospho-mTOR (S2448)-antibody 2 vials Stop Solution 1 x 8ml TMB One-Step Substrate Reagent 1 x 12ml -
Research areas
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Function
Kinase subunit of both mTORC1 and mTORC2, which regulates cell growth and survival in response to nutrient and hormonal signals. mTORC1 is activated in response to growth factors or amino-acids. Growth factor-stimulated mTORC1 activation involves AKT1-mediated phosphorylation of TSC1-TSC2, which leads to the activation of the RHEB GTPase that potently activates the protein kinase activity of mTORC1. Amino-acid-signaling to mTORC1 requires its relocalization to the lysosomes mediated by the Ragulator complex and the Rag GTPases. Activated mTORC1 up-regulates protein synthesis by phosphorylating key regulators of mRNA translation and ribosome synthesis. mTORC1 phosphorylates EIF4EBP1 and releases it from inhibiting the elongation initiation factor 4E (eiF4E). mTORC1 phosphorylates and activates S6K1 at 'Thr-421', which then promotes protein synthesis by phosphorylating PDCD4 and targeting it for degradation. Phosphorylates MAF1 leading to attenuation of its RNA polymerase III-repressive function. mTORC2 is also activated by growth. factors, but seems to be nutrient-insensitive. mTORC2 seems to function upstream of Rho GTPases to regulate the actin cytoskeleton, probably by activating one or more Rho-type guanine nucleotide exchange factors. mTORC2 promotes the serum-induced formation of stress-fibers or F-actin. mTORC2 plays a critical role in AKT1 'Ser-473' phosphorylation, which may facilitate the phosphorylation of the activation loop of AKT1 on 'Thr-308' by PDK1 which is a prerequisite for full activation. mTORC2 regulates the phosphorylation of SGK1 at 'Ser-422'. mTORC2 also modulates the phosphorylation of PRKCA on 'Ser-657'. -
Tissue specificity
Expressed in numerous tissues, with highest levels in testis. -
Sequence similarities
Belongs to the PI3/PI4-kinase family.
Contains 1 FAT domain.
Contains 1 FATC domain.
Contains 7 HEAT repeats.
Contains 1 PI3K/PI4K domain. -
Post-translational
modificationsAutophosphorylated; when part of mTORC1 or mTORC2. -
Cellular localization
Endoplasmic reticulum membrane. Golgi apparatus membrane. Mitochondrion outer membrane. Lysosome. Cytoplasm. Nucleus > PML body. Shuttles between cytoplasm and nucleus. Accumulates in the nucleus in response to hypoxia (By similarity). Targeting to lysosomes depends on amino acid availability and RRAGA and RRAGB. - Information by UniProt
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Alternative names
- dJ576K7.1 (FK506 binding protein 12 rapamycin associated protein 1)
- FK506 binding protein 12 rapamycin associated protein 1
- FK506 binding protein 12 rapamycin associated protein 2
see all -
Database links
- Entrez Gene: 2475 Human
- Omim: 601231 Human
- SwissProt: P42345 Human
- Unigene: 338207 Human
Images
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Positive Control
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.
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HEK-293 cells treated/untreated with FBS.HEK-293 cells were untreated or treated with 20% FBS for 20 minutes. -
HEK-293 cells treated/untreated with FBS.HEK-293 cells were untreated or treated with 20% FBS for 20 minutes.
Datasheets and documents
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SDS download
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Datasheet download
References (0)
ab279868 has not yet been referenced specifically in any publications.