Product Name:
RPTOR-pS859+pS863
Product Number:
ab-pn981
Target Full Name: Regulatory-associated protein of mTOR
Target Alias: KIAA1303; KOG1; Mip1; P150 target of rapamycin (TOR)-scaffold protein; RAPTOR; Regulatory associated protein of mTOR; Regulatory associated protein of MTOR, complex 1; Regulatory-associated protein of mTOR: P150 target of rapamycin (TOR)-scaffold protein; RPOR
Product Type Specific: RPTOR phosphosite-specific antibody
Antibody Code: PN981
Antibody Target Type: Phosphosite-specific
Antibody Phosphosite: S859+S863
Protein UniProt: Q8N122
Protein SigNET: RPTOR
Antibody Type: Polyclonal
Antibody Host Species: Rabbit
Antibody Immunogen Source: Synthetic phosphopeptide patterned after human RPTOR
Antibody Immunogen Sequence: LTQ(pS)APA(pS)PTN(βA)C
Antibody Immunogen Description: Corresponds to amino acid residues L856 to N866. The effects of Raptor S859 and S863 phosphorylation are unclear. These are major sites of in vivo phosphorylation of Raptor based on ≥74 and ≥107 high throughput mass spectrometry reports, respectively, recorded in PhosphoSitePlus. RPTOR is known to be phosphorylated in vitro at S859 by GSK3b, mTOR (FRAP1), and ULK1, and at S863 by ERK1 (MAPK3), ERK2 (MAPK1), GSK3b, JNK1 (MAPK8), mTOR (FRAP1), NLK, and p38b MAPK (MAPK11).
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.
Antibody Modification: Unconjugated. Contact KInexus if you are interest in having the antibody biotinylated or coupled with fluorescent dyes.
Antibody Concentration: 0.5 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 Species Reactivity: This antibody detects the target phosphoprotein in the following species due to conservation of amino acid sequence: Human | Chimpanzee | Dog | Rat | Mouse | Chicken | Frog | Zebra fish.
Related Product 1: RPTOR T706 phosphorylation antibody (Cat. No.: AB-PN982)
Scientific Background: RPTOR (Regulatory-associated protein of mTOR; Raptor) is a critical 150-kDa adaptor/scaffolding protein and essential component of the mTOR complex 1 (mTORC1). It belongs to the WD repeat RAPTOR family. It functions as a key sensor for nutrients such as amino acids, energy, and growth factor signaling. By promoting mTORC1 activity, RPTOR positively regulates protein synthesis, cell growth, and cell size while inhibiting autophagy. It features a unique N-terminal conserved (RNC) domain, three HEAT repeats, and seven WD40 repeats in the C-terminal region. As a scaffolding protein, it supports the structural integrity and localization of the mTORC1 complex to lysosomal membranes via Rag GTPases. It binds to TOS motif-containing proteins, directing substrates like 4E-BP1 and p70 S6 kinase to the active site of the mTOR kinase. Growth factor-stimulated mTORC1 activation involves a 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-389, which then promotes protein synthesis by phosphorylating PDCD4 and targeting it for degradation. Under energy stress, AMPK phosphorylates RPTOR, leading to binding with 14-3-3 proteins and subsequent inhibition of mTORC1, which helps trigger cell cycle arrest. It is involved in ciliogenesis. RPTOR is widely expressed, with high levels found in skeletal muscle, and to a lesser extent in brain, lung, small intestine, kidney, and placenta. Isoform 3 is widely expressed, with highest levels in nasal mucosa and pituitary and lowest in spleen. Its dysfunction is linked to improper nutrient sensing, cell growth defects, and various diseases. This description may include information annotated by UniProt and/or Google AI.