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mTOR Substrates Antibody Sampler Kit
mTOR Substrates Antibody Sampler Kit
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mTOR Substrates Antibody Sampler Kit

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    产品介绍
    产品信息
    抗原名称
    mTOR Substrates
    来源纯化
    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.
    简单描述
    Antibody Sampler Kit for studying in the research area.
    应用
    目标/特异性

    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.

    背景
    背景
    哺乳动物 rapamycin 靶标蛋白 (mTOR, FRAP, RAFT) 是一种 Ser/Thr 蛋白激酶 (1-3)，可作为ATP 和氨基酸的感受器来平衡营养可用性和细胞生长 (4,5)。当有足够的营养可用时，mTOR 响应磷脂酸介导的信号，将正向信号传送到 p70 S6 激酶，并参与 eIF4E 抑制因子 4E-BP1 的失活 (6)。这些活动可导致特定 mRNA 亚群的翻译。mTOR 通过 PI3 激酶/Akt 信号转导通路在 Ser2448 位点被磷酸化，并在 Ser2481 位点自磷酸化 (7,8)。mTOR 在细胞生长和稳态中发挥着重要作用，并且可能在肿瘤中受到异常调节。基于这些原因，人们目前正在将 mTOR 作为抗癌治疗的潜在靶标进行研究 (9)。mTOR (Raptor) 的调节相关蛋白与 mTOR 相互作用以介导 mTOR 信号转导至下游靶标 (10,11)。Raptor 通过其 TOR 信号转导 (TOS) 基序 与 mTOR 底物（例如 4E-BP1 和 p70 S6 激酶）结合，从而对 mTOR 介导的底物磷酸化必不可少 (12,13)。FKBP12-rapamycin 复合体与 mTOR 结合可抑制 mTOR-raptor 相互作用，这表明雷帕霉素具有抑制 mTOR 信号转导的机制 (14)。该 mTOR-raptor 相互作用以及其由营养和/或 rapamycin 进行的调节都依赖于被称为 GβL 的蛋白 (15)。GβL 是 mTOR 和 rictor（mTOR 的 rapamycin 不敏感伴侣）之间的雷帕霉素不敏感复合体的一部分，可能介导 rictor-mTOR 信号转导到 PKCα 和其他下游靶标 (16)。Rictor-mTOR 复合体已被确认为以前难以捉摸的 PDK2，可导致 Akt/PKB 在 Ser473 位点的磷酸化，这对 Akt/PKB 在 Thr308 位点的 PDK1 磷酸化和 Akt/PKB 的完全激活必不可少 (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.

    参考图片

    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.

    Western blot analysis of lysates from unsynchronized (U) and nocodazole (N) treated (50ng/ml for 48 hours) HT29 cells using Phospho-p70 S6 Kinase (Ser371) Antibody (B) and p70 S6 Kinase Antibody #9202 (D). Incubation of the nitrocellulose membrane with calf intestinal alkaline phosphatase (CIP) after Western transfer abolishes the phospho-p70 S6 Kinase signal (A), but has no effect on the total p70 S6 kinase signal (C).

    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 (10nM) #9904 or LY294002 (50uM) #9901, using Phospho-p70 S6 Kinase (Ser371) Antibody (upper) or p70 S6 Kinase Antibody #9202 (lower).

    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.

    Western blot analysis of extracts from serum starved or serum treated (20%) 293, NIH/3T3, and PC12 cells, using Phospho-p70 S6 Kinase (Thr389) (108D2) Rabbit mAb (upper), or p70 S6 Kinase (49D7) rabbit mAb #2708 (lower).

    Immunohistochemical analysis of paraffin-embedded human lymphoma using Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb.

    Immunohistochemical analysis using Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb on SignalSlide (TM) Phospho-Akt (Ser473) IHC Controls #8101 (paraffin-embedded LNCaP cells untreated (left) or LY294002-treated (right)).

    Flow cytometric analysis of Jurkat cells, untreated (green) or LY294002, Wortmannin and U0126-treated (blue), using Phospho-4E-BP1 (Thr36/46) (236B4) Rabbit mAb compared to a nonspecific negative control antibody (red).

    Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb.

    Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb in the presence of control peptide (left) or Phospho-4E-BP1 (Thr37/46) Blocking Peptide #1052 (right).

    Confocal immunofluorescent analysis of HeLa cells treated with LY294002 (left) or 20% serum (right) and labeled with Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).

    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分析不同细胞提取物，所用抗体为mTOR (7C10) Rabbit mAb兔单抗 #2983

    Western blot analysis of extracts from serum starved or serum treated (20%) 293, NIH/3T3, and PC12 cells, using Phospho-p70 S6 Kinase (Thr389) (108D2) Rabbit mAb #9234 (upper) or p70 S6 Kinase (49D7) Rabbit mAb #2708 (lower).

    Western blot 分析血清饥饿和20%血清处理的293, NIH/3T3, 和 PC12 细胞提取物，所用抗体为Phospho-p70 S6 Kinase (Thr389) (108D2) Rabbit mAb兔单抗 #9234 (上) 或 p70 S6 Kinase (49D7) Rabbit mAb 兔单抗#2708 (下)。

    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).western blot分析生长在低浓度血清中的293细胞，然后用20%的血清单独处理30分钟，或与雷帕霉素(10 nM) #9904 or LY294002 (50 uM) #9901预孵育1小时，所用抗体为Phospho-p70 S6 Kinase (Ser371) Antibody #9208 (上) 或 p70 S6 Kinase Antibody #9202 (下)

    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分析293T细胞提取物，所用抗体为4E-BP1 Antibody #9452 (u上) 和 Phospho-4E-BP1 (Thr37/46) Antibody #2855 (下). 细胞在无血清的培养液中饥饿24小时后，去除氨基酸1小时。在加入氨基酸1小时。未处理或采用100 nM insulin (+) 处理30分钟。

    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.

    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 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分析血清饥饿的NIH/3T3细胞，未处理组和胰岛素处理组 (150 nM, 5 分钟), 单独或与λ-磷酸酶联合组，所用抗体为Phospho-mTOR (Ser2448) (D9C2) XP® Rabbit mAb兔单抗 #5536 (上) 或 mTOR (7C10) Rabbit mAb兔单抗 #2983

    Immunohistochemical analysis of paraffin-embedded mouse brain using mTOR (7C10) Rabbit mAb.

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    货号：
    9862T
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