The antibody detects endogenous level of mTOR only when phosphorylated at serine 2448.
Purification
The antibody was affinity-purified from rabbit antiserum by affinity-chromatography usingepitope-specific phosphopeptide. The antibody against non-phosphopeptide was removedby chromatogramphy using non-phosphopeptide corresponding to the phosphorylation site.
Immunogen
Peptide sequence around phosphorylation site of pSer2448 (T-D-S (p) -Y-S) derived from Human mTOR. Antibodies were produced by immunizing rabbits with synthetic phosphopeptide and KLH conjugates.
Western blotting: 1:500-1:1000 Immunohistochemistry: 1:50-1:100 Immunofluorescence: 1:100-1:200
Restrictions
For Research Use only
Format
Liquid
Concentration
1 mg/mL
Buffer
Phosphate buffered saline (without Mg2+ and Ca2+), pH 7.4, 150 mM NaCl, 0.02 % sodium azide and 50 % glycerol.
Preservative
Sodium azide
Precaution of Use
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Storage
4 °C/-20 °C
Storage Comment
Store at -20 °C for long term preservation (recommended). Store at 4 °C for short term use.
Macias, Blazejczyk, Kazmierska, Caban, Skalecka, Tarkowski, Rodo, Konopacki, Jaworski: "Spatiotemporal characterization of mTOR kinase activity following kainic acid induced status epilepticus and analysis of rat brain response to chronic rapamycin treatment." in: PLoS ONE, Vol. 8, Issue 5, pp. e64455, (2013) (PubMed).
Ning, Huang, Dong, Sun, Zhang, Xu, Li: "5-Aza-2'-deoxycytidine inhibited PDGF-induced rat airway smooth muscle cell phenotypic switching." in: Archives of toxicology, Vol. 87, Issue 5, pp. 871-81, (2013) (PubMed).
Target
MTOR (mTOR)
(Mechanistic Target of Rapamycin (serine/threonine Kinase) (mTOR))
Kinase subunit of both mTORC1 and mTORC2, which regulate cell growth and survival in response to nutrient and hormonal signals. mTORC1 is activated in response to growth factors or amino-acids. Amino-acid-signaling to mTORC1 is mediated by Rag GTPases, which cause amino-acid-induced relocalization of mTOR within the endomembrane system. 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. 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. 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'.