Product Pathways - MAPK Signaling
Phospho-p90RSK (Ser380) (D5D8) Rabbit mAb #12032
|12032S||100 µl (10 western blots)||---||In Stock||---|
|12032||carrier free and custom formulation / quantity||email request|
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|W||1:1000||Human, Mouse, Rat, Monkey||Endogenous||90||Rabbit IgG|
Species cross-reactivity is determined by western blot.
Applications Key: W=Western Blotting, IP=Immunoprecipitation, F=Flow Cytometry
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).
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser377 of human p90RSK3 protein.
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.
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.
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.
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).
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).
- Fisher, T.L. and Blenis, J. (1996) Mol Cell Biol 16, 1212-9.
- Smith, J.A. et al. (1999) J Biol Chem 274, 2893-8.
- Dalby, K.N. et al. (1998) J Biol Chem 273, 1496-505.
- Roux, P.P. et al. (2003) Mol Cell Biol 23, 4796-804.
- Cargnello, M. and Roux, P.P. (2011) Microbiol Mol Biol Rev 75, 50-83.
- Romeo, Y. et al. (2012) Biochem J 441, 553-69.
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