| Applications: | CLIA |
| Reactivity: | Human |
| Note: | FOR SCIENTIFIC EDUCATIONAL RESEARCH USE ONLY (RUO). MUST NOT BE USED IN DIAGNOSTIC OR OTHER MEDICAL APPLICATIONS. |
| Sensitivity: | 18.75pg/mL |
| Detection Limit: | 31.25~2000pg/mL |
| Short Description : | This human MTOR kit is a highly sensitive in-vitro chemiluminescent immunoassay for the measurement of trace amounts of analytes in serum, plasma and other biological fluids. |
| Storage Instruction: | If unopened the kit may be stored at 2-8°C for up to 1 month. If the kit will not be used within 1 month, store the components separately, according to the component table in the manual. |
| Assay Time: | 3.5h |
| Detection: | Chemiluminescence |
| Gene Symbol: | MTOR |
| Gene ID: | 2475 |
| Uniprot ID: | MTOR_HUMAN |
| Specificity: | This kit recognizes Human mTOR in samples. No significant cross-reactivity or interference between Human mTOR and analogues was observed. |
| Sample Type: | Serum, plasma and other biological fluids |
| Tissue Specificity | Expressed in numerous tissues, with highest levels in testis. |
| Post Translational Modifications | Autophosphorylates when part of mTORC1 or mTORC2. Phosphorylation at Ser-1261, Ser-2159 and Thr-2164 promotes autophosphorylation. Phosphorylation in the kinase domain modulates the interactions of MTOR with RPTOR and AKT1S1/PRAS40 and leads to increased intrinsic mTORC1 kinase activity. Phosphorylation at Ser-2159 by TBK1 in response to growth factors and pathogen recognition receptors promotes mTORC1 activity. Phosphorylation at Thr-2173 in the ATP-binding region by AKT1 strongly reduces kinase activity. |
| Function | Serine/threonine protein kinase which is a central regulator of cellular metabolism, growth and survival in response to hormones, growth factors, nutrients, energy and stress signals. MTOR directly or indirectly regulates the phosphorylation of at least 800 proteins. Functions as part of 2 structurally and functionally distinct signaling complexes mTORC1 and mTORC2 (mTOR complex 1 and 2). In response to nutrients, growth factors or amino acids, mTORC1 is recruited to the lysosome membrane and promotes protein, lipid and nucleotide synthesis by phosphorylating key regulators of mRNA translation and ribosome synthesis. This includes phosphorylation of EIF4EBP1 and release of its inhibition toward the elongation initiation factor 4E (eiF4E). Moreover, phosphorylates and activates RPS6KB1 and RPS6KB2 that promote protein synthesis by modulating the activity of their downstream targets including ribosomal protein S6, eukaryotic translation initiation factor EIF4B, and the inhibitor of translation initiation PDCD4. Stimulates the pyrimidine biosynthesis pathway, both by acute regulation through RPS6KB1-mediated phosphorylation of the biosynthetic enzyme CAD, and delayed regulation, through transcriptional enhancement of the pentose phosphate pathway which produces 5-phosphoribosyl-1-pyrophosphate (PRPP), an allosteric activator of CAD at a later step in synthesis, this function is dependent on the mTORC1 complex. Regulates ribosome synthesis by activating RNA polymerase III-dependent transcription through phosphorylation and inhibition of MAF1 an RNA polymerase III-repressor. Activates dormant ribosomes by mediating phosphorylation of SERBP1, leading to SERBP1 inactivation and reactivation of translation. In parallel to protein synthesis, also regulates lipid synthesis through SREBF1/SREBP1 and LPIN1. To maintain energy homeostasis mTORC1 may also regulate mitochondrial biogenesis through regulation of PPARGC1A. In the same time, mTORC1 inhibits catabolic pathways: negatively regulates autophagy through phosphorylation of ULK1. Under nutrient sufficiency, phosphorylates ULK1 at 'Ser-758', disrupting the interaction with AMPK and preventing activation of ULK1. Also prevents autophagy through phosphorylation of the autophagy inhibitor DAP. Also prevents autophagy by phosphorylating RUBCNL/Pacer under nutrient-rich conditions. Prevents autophagy by mediating phosphorylation of AMBRA1, thereby inhibiting AMBRA1 ability to mediate ubiquitination of ULK1 and interaction between AMBRA1 and PPP2CA. mTORC1 exerts a feedback control on upstream growth factor signaling that includes phosphorylation and activation of GRB10 a INSR-dependent signaling suppressor. Among other potential targets mTORC1 may phosphorylate CLIP1 and regulate microtubules. The mTORC1 complex is inhibited in response to starvation and amino acid depletion. The non-canonical mTORC1 complex, which acts independently of RHEB, specifically mediates phosphorylation of MiT/TFE factors MITF, TFEB and TFE3 in the presence of nutrients, promoting their cytosolic retention and inactivation. Upon starvation or lysosomal stress, inhibition of mTORC1 induces dephosphorylation and nuclear translocation of TFEB and TFE3, promoting their transcription factor activity. The mTORC1 complex regulates pyroptosis in macrophages by promoting GSDMD oligomerization. MTOR phosphorylates RPTOR which in turn inhibits mTORC1. As part of the mTORC2 complex MTOR may regulate other cellular processes including survival and organization of the cytoskeleton. mTORC2 plays a critical role in the phosphorylation at 'Ser-473' of AKT1, a pro-survival effector of phosphoinositide 3-kinase, facilitating its activation by PDK1. mTORC2 may regulate the actin cytoskeleton, through phosphorylation of PRKCA, PXN and activation of the Rho-type guanine nucleotide exchange factors RHOA and RAC1A or RAC1B. mTORC2 also regulates the phosphorylation of SGK1 at 'Ser-422'. Regulates osteoclastogenesis by adjusting the expression of CEBPB isoforms. Plays an important regulatory role in the circadian clock function.regulates period length and rhythm amplitude of the suprachiasmatic nucleus (SCN) and liver clocks. |
| Protein Name | Serine/Threonine-Protein Kinase Mtor 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 |
| Database Links | Reactome: R-HSA-1257604 Reactome: R-HSA-1632852 Reactome: R-HSA-165159 Reactome: R-HSA-166208 Reactome: R-HSA-3371571 Reactome: R-HSA-380972 Reactome: R-HSA-389357 Reactome: R-HSA-5218920 Reactome: R-HSA-5628897 Reactome: R-HSA-5674400 Reactome: R-HSA-6804757 Reactome: R-HSA-8943724 Reactome: R-HSA-9639288 |
| Cellular Localisation | Lysosome Membrane Peripheral Membrane Protein Cytoplasmic Side Endoplasmic Reticulum Membrane Golgi Apparatus Membrane Mitochondrion Outer Membrane Cytoplasm Nucleus Pml Body Microsome Membrane Cytoplasmic Vesicle Phagosome Shuttles Between Cytoplasm And Nucleus Accumulates In The Nucleus In Response To Hypoxia Targeting To Lysosomes Depends On Amino Acid Availability And Rraga And Rragb Lysosome Targeting Also Depends On Interaction With Meak7 Translocates To The Lysosome Membrane In The Presence Of Tm4sf5 |
| Alternative CLIA Names | Serine/Threonine-Protein Kinase Mtor CLIA kit Fk506-Binding Protein 12-Rapamycin Complex-Associated Protein 1 CLIA kit Fkbp12-Rapamycin Complex-Associated Protein CLIA kit Mammalian Target Of Rapamycin CLIA kit Mtor CLIA kit Mechanistic Target Of Rapamycin CLIA kit Rapamycin And Fkbp12 Target 1 CLIA kit Rapamycin Target Protein 1 CLIA kit MTOR CLIA kit FRAP CLIA kit FRAP1 CLIA kit FRAP2 CLIA kit RAFT1 CLIA kit RAPT1 CLIA kit |
| Specificity | This kit recognizes Human mTOR in samples. No significant cross-reactivity or interference between Human mTOR and analogues was observed. |
| Reproducibility | Both intra-CV and inter-CV are |
Information sourced from Uniprot.org
| Item | Specifications | Storage |
|---|---|---|
| Micro CLIA Plate (Dismountable) | 96T: 8 wells ×12 strips strips | -20℃, 6 months |
| Reference Standard | 96T: 2 vials 48T: 1 vial | -20℃, 6 months |
| Concentrated Biotinylated Detection Ab (100×) | 96T: 1 vial, 120 μL 60 μL | -20℃, 6 months |
| Concentrated HRP Conjugate (100×) | 96T: 1 vial, 120 μL 60 μL | -20℃ (Protect from light), 6 months |
| Reference Standard & Sample Diluent | 1 vial, 20 mL | 2-8°C, 6 months |
| Biotinylated Detection Ab Diluent | 1 vial, 14 mL | 2-8°C, 6 months |
| HRP Conjugate Diluent | 1 vial, 14 mL | 2-8°C, 6 months |
| Concentrated Wash Buffer (25×) | 1 vial, 30 mL | 2-8°C, 6 months |
| Substrate Reagent A | 1 vial, 5 mL | 2-8℃ (Protect from light) |
| Substrate Reagent B | 1 vial, 5 mL | 2-8℃ (Protect from light) |
| Plate Sealer | 5 pieces | |
| Manual | 1 copy | |
| Certificate of Analysis | 1 copy |
| Sample Type | Range (%) | Average Recovery (%) |
|---|---|---|
| Serum(n=8) | 100-112 | 106 |
| EDTA plasma(n=8) | 94-105 | 100 |
| Cell culture media(n=8) | 92-104 | 99 |
| Intra-assay Precision | Intra-assay Precision | Intra-assay Precision | Inter-assay Precision | Inter-assay Precision | Inter-assay Precision | |
|---|---|---|---|---|---|---|
| Sample | 1.00 | 2.00 | 3.00 | 1.00 | 2.00 | 3.00 |
| n | 20.00 | 20.00 | 20.00 | 20.00 | 20.00 | 20.00 |
| Mean (pg/mL) | 96.68 | 278.01 | 899.95 | 87.25 | 253.37 | 883.59 |
| Standard deviation | 8.18 | 21.21 | 95.39 | 9.82 | 27.24 | 89.86 |
| CV (%) | 8.46 | 7.63 | 10.60 | 11.26 | 10.75 | 10.17 |