Cell Signaling Technology

Product Pathways - Protein Translation

Phospho-Drosophila p70 S6 Kinase (Thr398) Antibody #9209

70kda s6 kinase   dro-p70   drosophila   ds6k   ds6k1   p-ds6k1   p-s6k   p70-S6K   p70s6   s6k   s6kinase  

No. Size Price
9209S 100 µl ( 10 western blots ) ¥3,900.00 现货查询 购买询价
9209 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,D. melanogaster, Endogenous 70 Rabbit

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting,

Specificity / Sensitivity

Phospho-Drosophila p70 S6 Kinase (Thr398) Antibody detects endogenous levels of Drosophila p70 S6 kinase only when phosphorylated at threonine 398. The antibody will also recognize human p70 S6 Kinase when phosphorylated at threonine 389.

Phospho-Drosophila p70 S6 Kinase (Thr398) Antibody可以检测果蝇Thr389位点磷酸化的内源性p70 S6激酶。这些抗体也可以检测Thr389磷酸化的人的p70 S6激酶。

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr398 of Drosophila p70 S6 kinase. Antibodies are purified by protein A and peptide affinity chromatography.

单克隆抗体是采用果蝇p70 S6激酶Thr389位点周围残基相对应的合成磷酸肽免疫动物生产的,抗体采用蛋白A和肽亲和层析法纯化。

Western Blotting

Western Blotting

Western blot analysis of extracts from Drosophila S2 cells and Human MCF-7 cells, using Phospho-Drosophila p70 S6 Kinase (Thr398) Antibody (upper), Phospho-p70 S6 Kinase (Thr389) Antibody #9205, which recognizes the human, mouse and rat protein (middle), or Akt Antibody #9272 (lower). Serum-starved S2 cells were untreated or treated with EGF (100 ng/ml, 30min) #9908. Serum-starved MCF-7 cells were untreated or treated with IGF-1 (50 ng/ml) in the presence or absence of LY294002 (10 uM) #9901.

Western blot分析果蝇S2细胞和人MCF-7细胞提取物,所用抗体为Phospho-Drosophila p70 S6 Kinase (Thr398) Antibody (上), Phospho-p70 S6 Kinase (Thr389) Antibody #9205, 可以识别人类、小鼠和大鼠蛋白 (中) 或 Akt Antibody #9272 (下)。血清饥饿的S2细胞分为未处理组和用EGF (100 ng/ml, 30min) #9908处理组。血清饥饿的MCF-7细胞在LY294002 (10 uM) #9901存在和缺失的条件下,分为未处理组和采用IGF-1 (50 ng/ml)处理组。

Background

p70 S6 kinase is a mitogen activated Ser/Thr protein kinase that is required for cell growth and G1 cell cycle progression (1,2). p70 S6 kinase phosphorylates the S6 protein of the 40S ribosomal subunit and is involved in translational control of 5' oligopyrimidine tract mRNAs (1). A second isoform, p85 S6 kinase, is derived from the same gene and is identical to p70 S6 kinase except for 23 extra residues at the amino terminus, which encode a nuclear localizing signal (1). Both isoforms lie on a mitogen activated signaling pathway downstream of phosphoinositide-3 kinase (PI-3K) and the target of rapamycin, FRAP/mTOR, a pathway distinct from the Ras/MAP kinase cascade (1). The activity of p70 S6 kinase is controlled by multiple phosphorylation events located within the catalytic, linker and pseudosubstrate domains (1). Phosphorylation of Thr229 in the catalytic domain and Thr389 in the linker domain are most critical for kinase function (1). Phosphorylation of Thr389, however, most closely correlates with p70 kinase activity in vivo (3). Prior phosphorylation of Thr389 is required for the action of phosphoinositide 3-dependent protein kinase 1 (PDK1) on Thr229 (4,5). Phosphorylation of this site is stimulated by growth factors such as insulin, EGF and FGF, as well as by serum and some G-protein-coupled receptor ligands, and is blocked by wortmannin, LY294002 (PI-3K inhibitor) and rapamycin (FRAP/mTOR inhibitor) (1,6,7). Ser411, Thr421 and Ser424 lie within a Ser-Pro-rich region located in the pseudosubstrate region (1). Phosphorylation at these sites is thought to activate p70 S6 kinase via relief of pseudosubstrate suppression (1,2). Another LY294002 and rapamycin sensitive phosphorylation site, Ser371, is an in vitro substrate for mTOR and correlates well with the activity of a partially rapamycin resistant mutant p70 S6 kinase (8).Drosophila p70 S6 kinase (dS6K) is a critical regulator of cell growth, but this effect is independent of Akt and PI-3K signaling (9,10). However, insulin-induced activation and phosphorylation of dS6K at Thr398 is dependent on PI-3K, mTOR, and Akt (11,12).

p70 S6激酶是有丝分裂原活化的丝氨酸/苏氨酸蛋白激酶,是细胞生长和G1期细胞周期正常进行必需的(1,2)。P70 S6激酶可以磷酸化核糖体40S亚基的S6蛋白并参与5‘ oligopyrimidine tract mRNAs的翻译控制(1)。另一个亚型,P85 S6激酶,和p70 S6激酶由相同的基因转录,但氨基端有23个额外的残基,编码核定位信号(1)。两种亚型都处于有丝分裂原激活的PI-3K和rapamycin靶点FRAP/mTOR的信号通路下游,这条通路独立于Ras /MAP激酶级联反应(1)。p70 S6激酶活性取决于在催化、连接和伪底物结构域的多种磷酸化修饰(1)。催化结构域的Thr229和连接结构域的Thr389是调节激酶功能最关键的磷酸化修饰位点(1)。而且与P70激酶体内活性最相关的磷酸化修饰位点是Thr389(3)。Thr389磷酸化可以激活PDK1的Thr229磷酸化(4,5)。这个位点的磷酸化受到各种生长因子,如胰岛素,表皮生长因子和成纤维细胞生长因子,以及血清和一些G蛋白偶联受体配体,wortmannin,LY294002(PI-3K抑制剂)和rapamycin(FRAP/mTOR抑制剂)的刺激(1,6,7)。Ser411,Thr421和Thr424位于伪底物结构域的丝氨酸脯氨酸丰富区(1)。这些位点的磷酸化被认为是通过解除伪底物的抑制进而激活p70 S6激酶(1,2)。另一个对LY294002和rapamycin敏感的磷酸化位点,Ser371,是mTOR的体的底物,与某种抗rapamycin的P70 S6激酶突变体有关(8)。果蝇p70 S6激酶(dS6K)是细胞生长的重要调节因子,但这种效应依赖于Akt和PI-3K的信号(9,10)。然而,胰岛素诱导的dS6K在Thr398位点的活化和磷酸化依赖于PI-3K, mTOR, 和Akt (11,12)。

  1. Pullen, N. and Thomas, G. (1997) FEBS Lett 410, 78-82.
  2. Dufner, A. and Thomas, G. (1999) Exp Cell Res 253, 100-9.
  3. Weng, Q.P. et al. (1998) J Biol Chem 273, 16621-9.
  4. Pullen, N. et al. (1998) Science 279, 707-10.
  5. Alessi, D.R. et al. (1998) Curr Biol 8, 69-81.
  6. Polakiewicz, R.D. et al. (1998) J Biol Chem 273, 23534-41.
  7. Fingar, D.C. et al. (2002) Genes Dev 16, 1472-87.
  8. Saitoh, M. et al. (2002) J Biol Chem 277, 20104-12.
  9. Montagne, J. et al. (1999) Science 285, 2126-2129.
  10. Radimerski, T. et al. (2002) Nat. Cell Biol. 4, 251-255.
  11. Lizcano, J. M. et al. (2003) Biochem. J. 374, 297-306.
  12. Miron, M. et al. (2003) Mol. Cell. Biol. 23, 9117-9126.

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