GATOR Complex Antibody Sampler Kit #99732
Inquiry Info. # 99732
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Product Information
Kit Usage Information
Protocols
- 2983: Western Blotting, Immunoprecipitation (Magnetic), Immunohistochemistry (Paraffin), Immunofluorescence, Flow
- 5536: Western Blotting, Immunoprecipitation (Agarose), Immunofluorescence
- 7074: Western Blotting
- 13557: Western Blotting, Immunoprecipitation (Magnetic)
- 37344: Western Blotting, Immunoprecipitation (Agarose)
- 53385: Western Blotting, Immunoprecipitation (Agarose)
Product Description
The GATOR Complex Antibody Sampler Kit provides an economical means of detecting select components of the GATOR complex, mTOR and phospho-mTOR (Ser2448). The kit contains enough primary antibodies to perform at least two western blot experiments per antibody.
Specificity / Sensitivity
Mios (D12C6) Rabbit mAb recognizes endogenous levels of total Mios protein. NPRL2 (D8K3X) Rabbit mAb recognizes endogenous levels of total NPRL2 protein. WDR59 (D4Z7A) Rabbit mAb recognizes endogenous levels of total WDR59 protein. mTOR (7C10) Rabbit mAb detects endogenous levels of total mTOR protein. Phospho-mTOR (Ser2448) (D9C2) XP® Rabbit mAb detects endogenous levels of mTOR protein only when phosphorylated at Ser2448.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu730 of human Mios protein. Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala211 of human NPRL2 protein. Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding His356 of human WDR59 protein. Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser2481 of human mTOR. Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser2448 of human mTOR protein.
Background
The mTORC1 kinase complex plays a critical role in cell growth regulation (1,2). mTORC1 activity is modulated by energy levels, growth factors, and amino acids (3,4). Four related GTPases (RagA, RagB, RagC, and RagD) interact with raptor in mTORC1, which is necessary and sufficient for mTORC1 activation in response to amino acid signals (1,2). The GAP Activity Towards Rags (GATOR) complex interacts with Rag GTPases and is made up of a pair of protein subcomplexes (5). The GATOR1 subcomplex includes the proteins DEPDC5, Nprl2 and Nprl3, and is a RagA and RagB GTPase-activating protein (GAP) that negatively regulates mTORC1 signaling. Conversely, the GATOR2 subcomplex (including Mios, WDR24, WDR59, Seh1L and Sec13 proteins) is a positive regulator of mTORC1 signaling (5).
The mammalian target of rapamycin (mTOR, FRAP, RAFT) is a Ser/Thr protein kinase (6-8) that functions as an ATP and amino acid sensor to balance nutrient availability and cell growth (9,10). 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 (11). 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 (12,13).
The mammalian target of rapamycin (mTOR, FRAP, RAFT) is a Ser/Thr protein kinase (6-8) that functions as an ATP and amino acid sensor to balance nutrient availability and cell growth (9,10). 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 (11). 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 (12,13).
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- 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.
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