Revision 1
#89146
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For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:
W, IP
Reactivity:
H M R
Sensitivity:
Endogenous
MW (kDa):
150
Source/Isotype:
Rabbit IgG
UniProt ID:
#Q8N122
Entrez-Gene Id:
57521
Product Usage Information
Application | Dilution |
---|---|
Western Blotting | 1:1000 |
Immunoprecipitation | 1:50 |
Storage
Specificity/Sensitivity
Phospho-Raptor (Ser792) (E4V6C) Rabbit mAb recognizes endogenous levels of raptor protein only when phosphorylated at Ser792.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser792 of human raptor protein.
Background
The regulatory associated protein of mTOR (Raptor) was identified as an mTOR binding partner that mediates mTOR signaling to downstream targets (1,2). Raptor binds to mTOR substrates, including 4E-BP1 and p70 S6 kinase, through their TOR signaling (TOS) motifs and is required for mTOR-mediated phosphorylation of these substrates (3,4). Binding of the FKBP12-rapamycin complex to mTOR inhibits the mTOR-raptor interaction, suggesting a mechanism for rapamycin's specific inhibition of mTOR signaling (5). This mTOR-raptor interaction and its regulation by nutrients and/or rapamycin is dependent on a protein called GβL (6). GβL is also part of the rapamycin-insensitive complex between mTOR and rictor (rapamycin-insensitive companion of mTOR), and may mediate rictor-mTOR signaling to downstream targets including PKCα (7). Furthermore, the rictor-mTOR complex has been identified as the previously elusive PDK2 responsible for the phosphorylation of Akt/PKB on Ser473, facilitating phosphorylation of Akt/PKB on Thr308 by PDK1 and required for the full activation of Akt/PKB (8).
Recently raptor has been identified as a direct substrate of the AMP-activated protein kinase (AMPK) (9). AMPK phosphorylates raptor on Ser722/Ser792 (9). This phosphorylation is essential for inhibition of the raptor-containing mTOR complex 1 (mTORC1) and induces cell cycle arrest when cells are stressed for energy (9). These findings suggest that raptor is a critical switch that correlates cell cycle progression with energy status.
Background References
- Hara, K. et al. (2002) Cell 110, 177-89.
- Kim, D. et al. (2002) Cell 110, 163-75.
- Beugnet, A. et al. (2003) J. Biol. Chem. 278, 40717-22.
- Nojima, H. et al. (2003) J. Biol. Chem. 278, 15461-64.
- Oshiro, N. et al. (2004) Genes Cells 9, 359-66.
- Kim, D. H. et al. (2003) Mol. Cell 11, 895-904.
- Sarbassov, D. et al. (2004) Curr. Biol. 14, 1296-302.
- Sarbassov, D.D. et al. (2005) Science 307, 1098-101.
- Gwinn, D.M. et al. (2008) Mol Cell 30, 214-26.
Species Reactivity
Species reactivity is determined by testing in at least one approved application (e.g., western blot).
Western Blot Buffer
IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
Applications Key
W: Western Blotting IP: Immunoprecipitation
Cross-Reactivity Key
H: Human M: Mouse R: Rat Hm: Hamster Mk: Monkey Vir: Virus Mi: Mink C: Chicken Dm: D. melanogaster X: Xenopus Z: Zebrafish B: Bovine Dg: Dog Pg: Pig Sc: S. cerevisiae Ce: C. elegans Hr: Horse GP: Guinea Pig Rab: Rabbit G: Goat All: All Species Expected
Trademarks and Patents
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.
Limited Uses
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Revision 1
Western blot analysis of extracts from PANC-1 cells, vehicle-treated (-) or treated (+) with combinations of the following treatments as indicated: carbonyl cyanide m-chlorophenylhydrazone (CCCP) (100 μM, 2 hr) and λ phosphatase/calf intestinal alkaline phosphatase (CIP), using Phospho-Raptor (Ser792) (E4V6C) Rabbit mAb (upper) or Raptor (24C12) Rabbit mAb #2280 (lower).
Western blot analysis of extracts from C2C12 cells, vehicle-treated (-) or treated with carbonyl cyanide m-chlorophenylhydrazone (CCCP) (10 μg/mL, 30 min; +), using Phospho-Raptor (Ser792) (E4V6C) Rabbit mAb (upper) or Raptor (24C12) Rabbit mAb #2280 (lower).
Immunoprecipitation of phospho-raptor protein from PANC-1 extracts treated with carbonyl cyanide m-chlorophenylhydrazone (CCCP) (100 μM, 2 hr). Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Phospho-Raptor (Ser792) (E4V6C) Rabbit mAb. Western blot analysis was performed using Phospho-Raptor (Ser792) (E4V6C) Rabbit mAb. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used as the secondary antibody.