Product Pathways - Translational Control
mTOR Substrates Antibody Sampler Kit #9862
|Kit Includes||Quantity||Applications||Reactivity||MW (kDa)||Isotype|
|mTOR (7C10) Rabbit mAb #2983||40 µl||W IHC-P IF-IC F||H M R Mk (Hr)||289||Rabbit IgG|
|Phospho-p70 S6 Kinase (Thr389) (108D2) Rabbit mAb #9234||40 µl||W||H M R Mk (C)||70, 85||Rabbit IgG|
|Phospho-p70 S6 Kinase (Ser371) Antibody #9208||40 µl||W||H M R Mk||70, 85||Rabbit|
|Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb #2855||40 µl||W IHC-P IF-IC F||H M R Mk Dm||15 to 20||Rabbit IgG|
|Phospho-mTOR (Ser2448) (D9C2) XP® Rabbit mAb #5536||40 µl||W IP IF-IC||H M Mk (R) (C) (Pg) (Hr)||289||Rabbit IgG|
|Anti-rabbit IgG, HRP-linked Antibody #7074||100 µl||Goat|
Reactivity Key: H=Human M=Mouse R=Rat Mk=Monkey C=Chicken Dm=D. melanogaster Pg=Pig Hr=Horse
Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Specificity / Sensitivity
Each antibody in the mTOR Substrates Antibody Sampler Kit detects endogenous levels of its target protein. While activation state antibodies typically detect only target proteins phosphorylated at indicated residues, some cross-reaction can occur with related proteins phosphorylated at analogous sites.
Western blot analysis of extracts from 293T cells using 4E-BP1 Antibody #9452 (upper) and Phospho-4E-BP1 (Thr37/46) Antibody #2855 (lower). The cells were starved for 24 hours in serum-free medium and underwent a 1 hour amino acid deprivation. Amino acids were replenished for 1 hour. Cells were then either untreated (-) or treated with 100 nM insulin (+) for 30 minutes.
Western blot analysis of extracts from various cell types using mTOR (7C10) Rabbit mAb #2983.
Western blot analysis of extracts from serum-starved NIH/3T3 cells, untreated or insulin-treated (150 nM, 5 minutes), alone or in combination with λ-phosphatase, using Phospho-mTOR (Ser2448) (D9C2) XP® Rabbit mAb #5536 (upper) or mTOR (7C10) Rabbit mAb #2983.
Western blot analysis of lysates from 293 cells grown in low serum, then treated with 20% serum for 30 minutes alone or after 1 hour preincubation with rapamycin (10 nM) #9904 or LY294002 (50 uM) #9901, using Phospho-p70 S6 Kinase (Ser371) Antibody #9208 (upper) or p70 S6 Kinase Antibody #9202 (lower).
The mTOR Substrates Antibody Sampler Kit provides an economical means to evaluate the signaling of mTOR to downstream substrates including p70 S6 Kinase and 4E-BP1. The kit contains enough primary and secondary antibodies to perform four Western blot experiments per primary antibody.
Source / Purification
Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser371 of human p70 S6 kinase. Polyclonal antibodies are purified by protein A and peptide affinity chromatography. Phospho-specific rabbit monoclonal antibodies are produced by immunizing animals with synthetic phosphopeptides corresponding to residues surrounding Thr389 of human p70 S6 kinase, Thr37 and Thr46 of mouse 4E-BP1 and the Ser2448 site of human mTOR. The mTOR (7C10) Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser2481 of human mTOR.
The mammalian target of rapamycin (mTOR, FRAP, RAFT) is a Ser/Thr protein kinase (1-3) that functions as an ATP and amino acid sensor to balance nutrient availability and cell growth (4,5). When sufficient nutrients are available, mTOR responds to a phosphatidic acid-mediated signal to transmit a positive signal to p70 S6 kinase and participate in the inactivation of the eIF4E inhibitor, 4E-BP1 (6). These events result in the translation of specific mRNA subpopulations. mTOR is phosphorylated at Ser2448 via the PI3 kinase/Akt signaling pathway and autophosphorylated at Ser2481 (7,8). mTOR plays a key role in cell growth and homeostasis and may be abnormally regulated in tumors. For these reasons, mTOR is currently under investigation as a potential target for anti-cancer therapy (9).
The regulatory associated protein of mTOR (Raptor) interacts with mTOR to mediate mTOR signaling to downstream targets (10,11). Raptor binds to mTOR substrates, such as 4E-BP1 and p70 S6 kinase, through their TOR signaling (TOS) motifs and is required for mTOR-mediated substrate phosphorylation (12,13). Binding of the FKBP12-rapamycin complex to mTOR inhibits mTOR-raptor interaction, which suggests a mechanism for the inhibition of mTOR signaling by rapamycin (14). This mTOR-raptor interaction and its regulation by nutrients and/or rapamycin are dependent on a protein called GβL (15). GβL is part of the rapamycin-insensitive complex between mTOR and rictor (rapamycin-insensitive companion of mTOR) and may mediate rictor-mTOR signaling to PKCα and other downstream targets (16). The rictor-mTOR complex has been identified as the previously elusive PDK2 responsible for the phosphorylation of Akt/PKB at Ser473, which is required for PDK1 phosphorylation of Akt/PKB at Thr308 and full activation of Akt/PKB (17).
- Sabers, C.J. et al. (1995) J Biol Chem 270, 815-22.
- Brown, E.J. et al. (1994) Nature 369, 756-8.
- Sabatini, D.M. et al. (1994) Cell 78, 35-43.
- Gingras, A.C. et al. (2001) Genes Dev 15, 807-26.
- Dennis, P.B. et al. (2001) Science 294, 1102-5.
- Fang, Y. et al. (2001) Science 294, 1942-5.
- Navé, B.T. et al. (1999) Biochem J 344 Pt 2, 427-31.
- Peterson, R.T. et al. (2000) J Biol Chem 275, 7416-23.
- Huang, S. and Houghton, P.J. (2003) Curr Opin Pharmacol 3, 371-7.
- Hara, K. et al. (2002) Cell 110, 177-89.
- Kim, D.H. 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, 15461-4.
- Oshiro, N. et al. (2004) Genes Cells 9, 359-66.
- Kim, D.H. et al. (2003) Mol Cell 11, 895-904.
- Sarbassov, D.D. et al. (2004) Curr Biol 14, 1296-302.
- Sarbassov, D.D. et al. (2005) Science 307, 1098-101.
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- 9964 mTOR Pathway Antibody Sampler Kit
- 9864 mTOR Regulation Antibody Sampler Kit
- 2971 Phospho-mTOR (Ser2448) Antibody
- 2983 mTOR (7C10) Rabbit mAb
- 9234 Phospho-p70 S6 Kinase (Thr389) (108D2) Rabbit mAb
- 9208 Phospho-p70 S6 Kinase (Ser371) Antibody
- 2855 Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb
- 2974 Phospho-mTOR (Ser2481) Antibody
- 2972 mTOR Antibody
- 9205 Phospho-p70 S6 Kinase (Thr389) Antibody
- 9206 Phospho-p70 S6 Kinase (Thr389) (1A5) Mouse mAb
- 9459 Phospho-4E-BP1 (Thr37/46) Antibody
- 9456 Phospho-4E-BP1 (Ser65) (174A9) Rabbit mAb
- 9451 Phospho-4E-BP1 (Ser65) Antibody
- 9455 Phospho-4E-BP1 (Thr70) Antibody
- 2280 Raptor (24C12) Rabbit mAb
- 2114 Rictor (53A2) Rabbit mAb
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 7075 Anti-biotin, HRP-linked Antibody
- 7727 Biotinylated Protein Ladder Detection Pack
- 7003 20X LumiGLO® Reagent and 20X Peroxide
- 7071 Phototope®-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
For Research Use Only. Not For Use In Diagnostic Procedures.