Cell Signaling Technology

Product Pathways - MAPK Signaling

Phospho-p90RSK (Ser380) (D5D8) Rabbit mAb #12032

No. Size Price
12032S 100 µl ( 10 western blots ) ¥3,900.00 现货查询 购买询价
12032 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat,Monkey, Endogenous 90 Rabbit IgG
IP 1:100
F 1:400

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, F=Flow Cytometry,

Homology

Species predicted to react based on 100% sequence homology: Chicken, Xenopus, Zebrafish, Bovine, Dog, Pig, Horse,

Specificity / Sensitivity

Phospho-p90RSK (Ser380) (D5D8) Rabbit mAb recognizes endogenous levels of RSK1, RSK2, and RSK3 proteins only when phosphorylated at Ser380 (RSK1), Ser386 (RSK2), or Ser377 (RSK3).

Phospho-p90RSK (Ser380) (D5D8)Rabbit mAb兔单抗可以识别内源性的Ser380 (RSK1), Ser386 (RSK2)或 Ser377 (RSK3)被磷酸化后的RSK1, RSK2及RSK3蛋白。

Source / Purification

Monoclonal antibody is produced by immunizing animals with synthetic peptides corresponding to residues surrounding Ser377 of human RSK3 protein and Ser386 of RSK2 protein.

单克隆抗体由合成肽段免疫动物产生,该肽段与人RSK3的Ser377以及RSK2 的Ser386及其邻近氨基酸序列一致。

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines, starved overnight and untreated (-) or treated with either TPA #4174 (200 nM, 15 min; +), hEGF #8916 (100 ng/ml, 15 min; +), or hPDGF-BB #8912 (100 ng/ml, 15 min; +), using Phospho-p90RSK (Ser380) (D5D8) Rabbit mAb.血清饥饿过夜然后不处理(-)或用TPA #4174 (200 nM, 15 min)及Human hEGF #8916 (100 ng/mL, 15 min)处理(+),或TPA #4174 (200 nM, 15 min)及hPDGF-BB #8912 (100 ng/ml, 15 min; +)处理的多种细胞提取物进行,使用Phospho-p90RSK (Ser380) (D5D8)Rabbit mAb进行western blot分析。

IP

IP

Immunoprecipitation of p90RSK from HeLa cell extracts, serum starved overnight and untreated (-) or treated with TPA #4174 (200 nM, 15 min; +), using Normal Rabbit IgG #2729 (lanes 5 and 6) or Phospho-p90RSK (Ser380) (D5D8) Rabbit mAb (lanes 3 and 4). Lanes 1 and 2 are 10% input. Western blot analysis was performed using Phospho-p90RSK (Ser380) Antibody #9341.血清饥饿过夜后不处理(-)或使用TPA #4174 (200 nM, 15 min; +)处理的HeLa细胞提取物,使用Rabbit IgG #2729 (lanes 5和 6)或Phospho-p90RSK (Ser380) (D5D8) 兔mAb(lanes 3和 4)对 p90RSK进行免疫沉淀实验。Lanes1和2是10%上样量。Phospho-p90RSK (Ser380)抗体#9341用作western blot分析。

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells, untreated (blue) or treated with TPA #4174 (green), using Phospho-p90RSK (Ser380) (D5D8) Rabbit mAb. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 647 Conjugate) #4414 was used as a secondary antibody.未处理(蓝色)或经过TPA #4174(绿色)处理的Jurkat细胞,使用Phospho-p90RSK (Ser380) (D5D8)Rabbit mAb进行流式分析。Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 647 Conjugate) #4414作为二抗。

Background

The 90 kDa ribosomal S6 kinases (RSK1-4) are a family of widely expressed Ser/Thr kinases characterized by two nonidentical, functional kinase domains (1) and a carboxy-terminal docking site for extracellular signal-regulated kinases (ERKs) (2). Several sites both within and outside of the RSK kinase domain, including Ser380, Thr359, Ser363, and Thr573, are important for kinase activation (3). RSK1-3 are activated via coordinated phosphorylation by MAPKs, autophosphorylation, and phosphoinositide-3-OH kinase (PI3K) in response to many growth factors, polypeptide hormones, and neurotransmitters (3).

90kDa核糖体S6激酶(RSK1-4)是一个广泛表达的丝/苏氨酸激酶家族,其特征是包含2个不同的功能性激酶结构域(1)和一个羧基端的细胞外信号调节激酶(ERKs)结合位点(2)。RSK激酶结构域内或以外的一些位点,包括Ser380, Thr359, Ser363和Thr573,对于激酶的激活非常重要(3)。RSK1-3在多种生长因子、多肽激素和神经递质的刺激下,通过MAPKs的磷酸化作用、自磷酸化作用以及PI3K的信号途径的协同作用被激活(3)。

Upon mitogenic stimulation, p44/42 ERK1/2 and ERK5 MAP kinases cooperatively phosphorylate p90RSK Thr573 (p90RSK1 numbering) located within the C-terminal kinase domain and Thr359/Ser363 in the linker region between the two kinase domains (3). Phosphorylation of Thr573 within the activation loop of the p90RSK C-terminal kinase domain promotes activation and directs phosphorylation of Ser380 within the hydrophobic stretch of the linker region (4,5). The phosphorylated p90RSK Ser380 acts as a docking site for the constitutively active Ser/Thr kinase PDK1, which in turn phosphorylates Ser221 within the N-terminal kinase domain activation loop, resulting in full enzymatic activation of the p90RSK (6). Antibodies against these phosphorylation sites are useful for understanding the kinetics and regulation of p90RSK activation. 
 For more information regarding the phospho-regulatory sites within each RSK isoform, including more information regarding the seminal studies demonstrating the complex phosphorylation cascades involved, please see the references herein and PhosphoSitePlus® (www.phosphosite.org).

在有丝分裂原的刺激下,p44/42 Erk1/2和Erk5 MAPK能够协同磷酸化p90RSK位于C末端激酶结构域的Thr573 (p90RSK1 numbering)以及位于两个激酶结构域间链接序列的Thr359/Ser363 (3)。p90RSK C末端激酶结构域活化环内Thr573位点的磷酸化能够促进连接区域疏水区内的Ser380位点的磷酸化及活化过程(4,5)。被磷酸化后,Ser380能够作为组成型活化的Ser/Thr激酶PDK1的结合位点,随后使p90RSK的N末端激酶结构活化环内的Ser221位点被磷酸化,导致p90RSK酶活性的全面激活(6)。针对这些磷酸化位点的抗体对于理解p90RSK活性的动力学原理和调控是有用的。为了获得更多每个RSK亚型磷酸化位点的信息,包括证明这些复合物磷酸化所涉及的级联反应,请参考参考文献和PhosphoSitePlus® (www.phosphosite.org)。

  1. Fisher, T.L. and Blenis, J. (1996) Mol Cell Biol 16, 1212-9.
  2. Smith, J.A. et al. (1999) J Biol Chem 274, 2893-8.
  3. Dalby, K.N. et al. (1998) J Biol Chem 273, 1496-505.
  4. Roux, P.P. et al. (2003) Mol Cell Biol 23, 4796-804.
  5. Cargnello, M. and Roux, P.P. (2011) Microbiol Mol Biol Rev 75, 50-83.
  6. Romeo, Y. et al. (2012) Biochem J 441, 553-69.

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