mTOR Regulation Antibody Sampler Kit

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mTOR Regulation Antibody Sampler Kit

品牌： CST

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产品介绍

产品信息

抗原名称

mTOR Regulation

来源纯化

Phospho-specific polyclonal antibody is produced by immunizing animals with synthetic phosphopeptides corresponding to residues surrounding Ser792 of human raptor. Polyclonal antibodies are purified by protein A and peptide affinity chromatography. Phospho-specific monoclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to the sequence surrounding Thr246 of human PRAS40 and Ser2448 of human mTOR. Total protein monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Ser2481 of human mTOR, the sequence of human PRAS40, and the residues near the aminoterminus of human RagC.

产品详情

PI3K / Akt Signaling

简单描述

Antibody Sampler Kit for studying mTOR/mTOR (Ser2448) phosphate/Raptor (Ser792) phosphate/RagC/PRAS40 (Thr246) phosphate/PRAS40 in the research area.

研究领域

癌症,细胞生物学,纤维化,代谢,神经科学

应用

目标/特异性

Specificity/Sensitivity

Each antibody in the mTOR Regulation Antibody Sampler Kit detects endogenous levels of its target protein. Activation state antibodies detect only target proteins phosphorylated at indicated residues. Phospho-Raptor (Ser792) Antibody may also detect non-specific signals of various molecular weights.

背景

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) was identified as an mTOR binding partner that mediates mTOR signaling to downstream targets (10,11). 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 (12,13). PRAS40 interacts with raptor in insulin-deprived cells and inhibits the activation of the mTORC1 pathway. Phosphorylation of PRAS40 by Akt at Thr246 relieves PRAS40 inhibition of mTORC1 (14). Recently raptor has been identified as a direct substrate of the AMP-activated protein kinase (AMPK) (15). AMPK phosphorylates raptor on Ser722/Ser792 (15). 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 (15). These findings suggest that raptor is a critical switch that correlates cell cycle progression with energy status. The activity of mTORC1 kinase complex is modulated by energy levels, growth factors and amino acids (16,17). Recent studies found that RagA, RagB, RagC and RagD, the four related GTPases, interact with raptor in the mTORC1 complex (18,19). These interactions are both necessary and sufficient for mTORC1 activation in response to amino acid signals (18,19). 1.Sabers, C.J. et al. (1995) J Biol Chem 270, 815-22. 2.Brown, E.J. et al. (1994) Nature 369, 756-8. 3.Sabatini, D.M. et al. (1994) Cell 78, 35-43. 4.Gingras, A.C. et al. (2001) Genes Dev 15, 807-26. 5.Dennis, P.B. et al. (2001) Science 294, 1102-5. 6.Fang, Y. et al. (2001) Science 294, 1942-5. 7.Navé, B.T. et al. (1999) Biochem J 344 Pt 2, 427-31. 8.Peterson, R.T. et al. (2000) J Biol Chem 275, 7416-23. 9.Huang, S. and Houghton, P.J. (2003) Curr Opin Pharmacol 3, 371-7. 10.Hara, K. et al. (2002) Cell 110, 177-89. 11.Kim, D.H. et al. (2002) Cell 110, 163-75. 12.Beugnet, A. et al. (2003) J Biol Chem 278, 40717-22. 13.Nojima, H. et al. (2003) J Biol Chem 278, 15461-4. 14.Vander Haar, E. et al. (2007) Nat Cell Biol 9, 316-23. 15.Gwinn, D.M. et al. (2008) Mol Cell 30, 214-26. 16.Hay, N. and Sonenberg, N. (2004) Genes Dev 18, 1926-45. 17.Wullschleger, S. et al. (2006) Cell 124, 471-84. 18.Sancak, Y. et al. (2008) Science 320, 1496-501. 19.Kim, E. et al. (2008) Nat Cell Biol 10, 935-45.

研究领域

癌症,细胞生物学,纤维化,代谢,神经科学

数据库链接

Entrez-Gene ID

2475,57521,64121,84335

UniProt ID

P42345,Q8N122,Q9HB90,Q96B36

参考图片

Immunohistochemical analysis of paraffin-embedded human lymphoma using RagC (D31G9) XP^® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).

After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO* is added and emits light during enzyme catalyzed decomposition.

Flow cytometric analysis of 293 cells using mTOR (7C10) Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, showing cytoplasmic localization using mTOR (7C10) Rabbit mAb.

Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using mTOR (7C10) Rabbit mAb in the presence of control peptide (left) or mTOR Blocking Peptide #1072 (right).

Western blot analysis of extracts from 293, A431, COS, C6, and C2C12 cells, using mTOR (7C10) Rabbit mAb.

Confocal immunofluorescent analysis of mouse embryonic fibroblast (MEF) cells using mTOR (7C10) Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5^® #4084 (fluorescent DNA dye).

Western blot analysis of wild-type (WT) and AMPKα1 and α2 knockout (KO) mouse embryonic fibroblasts (MEFs), untreated or treated with AICAR (2 mM for 1 hour), using Phospho-Raptor (Ser792) Antibody (upper) or Raptor Antibody #4978 (lower). (Image provided by Dr. Reuben Shaw, Salk Institute for Biological Studies).

*Cross-reacting bands at 60, 70 and 240 kDa

Western blot analysis of C2C12 or 293 cells, untreated or treated with AICAR (0.5 mM for 30 minutes) or oligomycin (0.5 μM for 30 minutes), using Phospho-Raptor (Ser792) Antibody (upper and lower left ) or Raptor Antibody #2280 (upper and lower right).

*Cross-reacting bands at 200 kDa.

Western blot analysis of extracts from various cell types using PRAS40 (D23C7) XP^® Rabbit mAb.

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using PRAS40 (D23C7) XP^® Rabbit mAb.

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, control (left) or λ phosphatase-treated (right), using Phospho-PRAS40 (Thr246) (C77D7) Rabbit mAb.

Immunohistochemical analysis of paraffin-embedded metastatic SKOV-3 tumor in mouse lung using Phospho-PRAS40 (Thr246) (C77D7) Rabbit mAb.

Western blot analysis of extracts from serum starved H3255, Mkn45 and NIH/3T3 cells, untreated or treated with either Iressa (1 μM, 3 hours), Su11274 (1 μM, 3 hours) or insulin (150 nM, 15 minutes), using Phospho-PRAS40 (Thr246) (C77D7) Rabbit mAb (upper) or PRAS40 (D23C7) Rabbit mAb #2691 (lower).

Western blot analysis of extracts from serum starved HeLa cells, untreated or treated with insulin (100 nM, 5 minutes) or with insulin and λ phosphatase, using Phospho-PRAS40 (Thr246) (C77D7) Rabbit mAb (upper) or PRAS40 Antibody #2610 (lower).

Western blot analysis of extracts from various cell types using mTOR (7C10) Rabbit mAb #2983.western blot 分析不同细胞系提取物，所用抗体为mTOR (7C10) Rabbit mAb #2983。

Western blot analysis of C2C12 or 293 cells, untreated or treated with AICAR (0.5 mM for 30 minutes) or oligomycin (0.5 μM for 30 minutes), using Phospho-Raptor (Ser792) Antibody #2083 (upper and lower left) or Raptor Antibody #2280 (upper and lower right). *Cross-reacting bands at 200 kDa.

Western blot分析C2C12 或293 cells，未处理组或AICAR (0.5 mM for 30 分钟) 或 oligomycin (0.5 μM for 30 分钟)处理组，所用抗体为Phospho-Raptor (Ser792) Antibody #2083 (上和左下图) 或 Raptor Antibody #2280 (上和右下图). *与200 kDa的条带发生交叉反应。

Western blot analysis of extracts from serum starved H3255, Mkn45 and NIH/3T3 cells, untreated or treated with either Iressa (1 μM, 3 hours), Su11274 (1 μM, 3 hours) or insulin (150 nM, 15 minutes), using Phospho-PRAS40 (Thr246) (C77D7) Rabbit mAb #2997 (upper) or PRAS40 (D23C7) Rabbit mAb #2691 (lower).

western blot 分析血清饥饿的H3255, Mkn45 和NIH/3T3细胞提取物，未处理组和Iressa (1 μM, 3 小时), Su11274 (1 μM, 3 小时)或胰岛素 (150 nM, 15 分钟)处理组。所用抗体为 Phospho-PRAS40 (Thr246) (C77D7) Rabbit mAb #2997兔单抗 (上) 或 PRAS40 (D23C7) Rabbit mAb兔单抗 #2691 (下)

Western blot analysis of extracts from various cell types using PRAS40 (D23C7) XP® Rabbit mAb #2691.

western blot 分析不同细胞系提取物，所用抗体为PRAS40 (D23C7) XP® Rabbit mAb 兔单抗#2691.

Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-) or SignalSilence^® mTOR siRNA II (+), using mTOR (7C10) Rabbit mAb #2983 and α-Tubulin (11H10) Rabbit mAb #2125. mTOR (7C10) Rabbit mAb confirms silencing of mTOR expression, while the α-Tubulin (11H10) Rabbit mAb is used to control for loading and specificity of mTOR siRNA.

Confocal immunofluorescent analysis of HeLa cells, rapamycin-treated (#9904, 10 μM for 2 hours, left), insulin-treated (150 nM for 6 minutes, middle) or insulin- and λ-phosphatase-treated (right), using Phospho-mTOR (Ser2448) (D9C2) XP^® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin. Blue pseudocolor = DRAQ5^® #4084 (fluorescent DNA dye).

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 (upper) or mTOR (7C10) Rabbit mAb #2983.

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using RagC (D31G9) XP^® Rabbit mAb.

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9864T

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