Product Pathways - PathScan Multiplex WB Cocktails
PathScan® Multiplex Western Cocktail II: Phospho-p90RSK, Phospho-p53, Phospho-p38 MAPK and Phospho-S6 Ribosomal Protein Detection Cocktail II #5302
|5302S||250 µl (5 western blots)||---||In Stock||---|
|5302||carrier free and custom formulation / quantity||email request|
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|W||1:200||Human, Mouse, Rat, Mink||Endogenous||Rabbit|
Species cross-reactivity is determined by western blot.
Applications Key: W=Western Blotting
Antibodies In This Cocktail
- Phospho-p90RSK (Ser380) Antibody #9341
- Phospho-p53 (Ser15) Antibody #9284
- Phospho-p38 MAP Kinase (Thr180/Tyr182) Antibody #9211
- Phospho-S6 Ribosomal Protein (Ser235/236) Antibody #2211
- eIF4E Antibody #9742
The PathScan® Multiplex Western Cocktail II offers a unique method to assay the activation of multiple pathways on one membrane without stripping and reprobing. This method saves the user valuable time, while increasing accuracy and minimizing reagent waste. The system allows the user to simultaneously detect levels of phospho-p90RSK, phospho-p53, phospho-p38 MAPK and phospho-S6 ribosomal protein. The kit also includes elF4E antibody to control protein loading.
Specificity / Sensitivity
Each phospho-antibody in this cocktail recognizes endogenous levels of only the phosphorylated form of its specific target. The eIF4E antibody detects endogenous levels of its target protein independent of phosphorylation and is provided to control for protein loading.
Source / Purification
Polyclonal antibodies are produced by immunizing animals with synthetic peptides. Antibodies are purified by protein A and peptide affinity chromatography.
The 90 kDa ribosomal S6 kinases (RSK1-3) are a family of serine/threonine kinases broadly expressed in response to many growth factors, polypeptide hormones and neurotransmitters (1). p90RSK is activated by Erk1 and Erk2 in vitro and in vivo via phosphorylation (2). Several sites, such as Ser380, Thr359 and Ser363, are important for its activation (3).
The p53 tumor suppressor protein plays a major role in cellular response to DNA damage and other genomic aberrations. Activation of p53 can lead to either cell cycle arrest and DNA repair or apoptosis (4). p53 is phosphorylated at multiple sites in vivo and by several different protein kinases in vitro (5,6). DNA damage induces phosphorylation of p53 at Ser15 and Ser20 and leads to reduced interaction of p53 with its negative regulator, oncoprotein MDM2 (7).
p38 MAP kinase controls cellular responses to cytokines and stress (8-11). Like the SAPK/JNK pathway, p38 MAP kinase is activated by a variety of cellular stresses including osmotic shock, inflammatory cytokines, lipopolysaccharides (LPS), UV light and growth factors (8-12). MKK3, MKK6 and SEK activate p38 MAP kinase by phosphorylation at Thr180 and Tyr182.
Growth factors and mitogens induce the activation of p70 S6 kinase, which in turn phosphorylates the S6 ribosomal protein. Phosphorylation of S6 correlates with an increase in translation, particularly of mRNAs with an oligopyrimidine tract in their 5' untranslated regions (13). This group of mRNAs (5'TOP) encodes proteins involved in cell cycle progression and proteins that are part of the translational machinery, such as ribosomal proteins and elongation factors (13,14).
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- Milczarek, G.J. et al. (1997) Life Sci. 60, 1-11.
- Shieh, S.Y. et al. (1997) Cell 91, 325-334.
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- Rouse, J. et al. (1994) Cell 78, 1027-1037.
- Freshney, N.W. et al. (1994) Cell 78, 1039-1049.
- Raingeaud, J. et al. (1995) J. Biol. Chem. 270, 7420-7426.
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- 2211 Phospho-S6 Ribosomal Protein (Ser235/236) Antibody
- 9211 Phospho-p38 MAPK (Thr180/Tyr182) Antibody
- 9284 Phospho-p53 (Ser15) Antibody
- 9341 Phospho-p90RSK (Ser380) Antibody
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 7071 Phototope®-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
- 7727 Biotinylated Protein Ladder Detection Pack
- 7003 20X LumiGLO® Reagent and 20X Peroxide
For Research Use Only. Not For Use In Diagnostic Procedures.
PathScan® is a trademark of Cell Signaling Technology, Inc.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.