Product Name:
mTOR-pS2478+pS2481
Product Number:
ab-pk730
Target Full Name: Mammalian target of rapamycin (FRAP); FKBP12-rapamycin complex-associated protein
Target Alias: DJ576K7.1; FK506 binding protein 12-rapamycin associated protein 1; FK506 binding protein 12-rapamycin associated protein 2; FK506-binding protein 12-rapamycin complex-associated protein 1; FKBP12-rapamycin complex-associated protein 1; FKBP-rapamycin associated protein; FLJ44809; FRAP; FRAP2; RAPT1; mTOR; CCDS127.1; ENSG00000198793
Product Type Specific: mTOR protein kinase phosphosite-specific antibody
Antibody Code: PK730
Antibody Target Type: Phosphosite-specific
Antibody Phosphosite: S2478+S2481
Protein UniProt: P42345
Protein SigNET: mTOR
Antibody Type: Polyclonal
Antibody Host Species: Rabbit
Antibody Immunogen Source: Human mTOR (FRAP) sequence peptide Cat. No.: PE-04AMM95
Antibody Immunogen Sequence: VPE(pS)IH(pS)FIG(bA)C
Antibody Immunogen Description: Corresponds to amino acid residues V2475 to G2484; in the C-terminal fifth of the kinase at the end of the PI3_PI4-kinase domain.S2448 phosphorylation stimulates phosphotransferase activity. This is the major in vivo phosphorylation site in mTOR (≥114 reports from high throughput mass spectrometry studies recorded in PhosphoSitePlus). mTOR is known to be phosphorylated at this site in vitro by Akt1 (PKBa), and p70S6K (RPS6KB1).
Production Method: The immunizing peptide was produced by solid phase synthesis on a multipep peptide synthesizer and purified by reverse-phase hplc chromatography. Purity was assessed by analytical hplc and the amino acid sequence confirmed by mass spectrometry analysis. This peptide was coupled to KLH prior to immunization into rabbits. New Zealand White rabbits were subcutaneously injected with KLH-coupled immunizing peptide every 4 weeks for 4 months. The sera from these animals was applied onto an agarose column to which the immunogen peptide was thio-linked. Antibody was eluted from the column with 0.1 M glycine, pH 2.5. Subsequently, the antibody solution was neutralized to pH 7.0 with saturated Tris.This antibody was also subject to negative purification over phosphotyrosine-agarose.
Antibody Modification: Unconjugated. Contact KInexus if you are interest in having the antibody biotinylated or coupled with fluorescent dyes.
Antibody Concentration: 0.75 mg/ml
Storage Buffer: Phosphate buffered saline pH 7.4, 0.05% Thimerasol
Storage Conditions: For long term storage, keep frozen at -40°C or lower. Stock solution can be kept at +4°C for more than 3 months. Avoid repeated freeze-thaw cycles.
Product Use: Western blotting | Antibody microarray
Antibody Dilution Recommended: 2 µg/ml for immunoblotting
Antibody Potency: Very strong immunoreactivity with immunogen peptide on dot blots.
Antibody Species Reactivity: This antibody detects the target phosphoprotein in the following species due to conservation of amino acid sequence: Human | Dog | Rat | Mouse | Platypus | Chicken | Zebra fish.
Antibody Positive Control: The observed molecular mass of the processed target protein on SDS-PAGE gels is reported to be around 250-300 kDa.
Antibody Specificity: Very high
Antibody Cross Reactivity: No significant cross-reactive proteins detected in MCF7 and T98G cells.
Related Product 1: mTORSubtide kinase substrate peptide (Cat. No.: PE-01BIJ90)
Related Product 3: mTOR expression antibody (Cat. No.: AB-NK116-5)
Related Product 4: mTOR expression antibody (Cat. No.: AB-NK116-4)
Related Product 5: mTOR expression antibody (Cat. No.: AB-NK116-3)
Related Product 6: mTOR T190 phosphorylation antibody (Cat. No.: AB-PK729)
Scientific Background: mTOR (FRAP1) is a 289 kDa protein-serine/threonine kinase of the Atypical group and PIKK family. It mediates cell growth, proliferation, metabolism, and survival. It senses nutrient, energy, and growth factor levels to balance cell metabolism. mTOR exists in two distinct complexes, mTORC1 (growth/protein synthesis) and mTORC2 (cytoskeleton/survival). mTORC1 integrates signals from growth factors like insulin, energy status (via AMPK), and amino acids (leucine, arginine) to regulate protein synthesis and inhibit autophagy. mTORC2 regulates the actin cytoskeleton, cell polarity and migration, and metabolism, including Akt/PKB phosphorylation. It targets eIF4e and p70S6 kinase activation for protein synthesis. Phosphorylation of Ser-2448 and Ser-2481 increases its phosphotransferase activity, and it is stimulated by RHEBm. Phosphorylation of Thr-2446 inhibits phosphorylation of the Ser-2448 activatory site. It is also negatively regulated by DEPDC6. mTOR has several functional domains: FAT Domain (Amino acids 1382-1982); PI3K/PI4K Catalytic Domain (Amino acids 2156-2469); FATC Domain (Amino acids 2517-2549); an interaction region for binding to FKBP1A/rapamycin (Amino acids 2012-2144); and an NBN interaction region (Amino acids 1-651). Dysregulation in signalling that leads to constant activation of mTOR lead to uncontrolled proliferation and cell cycle progression and oncogensis. Increased invasiveness has also be shown to be associated with mTOR activity. While it features a wide range of mutations, this might reflect the very large size of the protein rather than its identification as a tumour suppressor protein. Cancer-related mutations for mTOR in human tumours point to a gain of function of the protein kinase, indicating that it might function as an oncoprotein (OP). The active form of the protein kinase normally acts to promote tumour cell proliferation. Defective mTOR signalling has been linked with diabetes, Tuberous Sclerosis, Subependymal Giant Cell Astrocytoma (SEGA), Lymphangioleiomyomatosis (LAM), Tuberous Sclerosis Complex, Plasmablastic Lymphoma (PBL), Ewing's tumours, and Kidney Angiomyolipoma. Rapamycin and its analogs (rapalogs) inhibit mTOR, and are useful in cancer treatment. This description may include information annotated by UniProt and/or Google AI.