Cell Signaling Technology

Product Pathways - MAPK Signaling

Phospho-p38 MAPK (Thr180/Tyr182) (3D7) Rabbit mAb #9215

Applications Reactivity Sensitivity MW (kDa) Isotype
W IF-IC F H M R Mk Dm Pg Sc (Hm) (B) Endogenous 43 Rabbit IgG

Applications Key:  W=Western Blotting  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  Hm=Hamster  Mk=Monkey  Dm=D. melanogaster  B=Bovine  Pg=Pig  Sc=S. cerevisiae
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Phospho-p38 MAP Kinase (Thr180/Tyr182) (3D7) Rabbit mAb detects endogenous levels of p38 MAPK only when dually phosphorylated at Thr180 and Tyr182. This antibody does not cross-react with the phosphorylated forms of either p42/44 MAPK or SAPK/JNK.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr180/Tyr182 of human p38 MAPK.

Western Blotting

Western Blotting

Specificity of Phospho-Erk1/2, Phospho-p38 MAPK and Phospho-SAPK/JNK Rabbit mAb: Western blot analysis of extracts from NIH/3T3 cells treated with PDGF and UV, using Phospho-p38 MAPK Rabbit mAb #9215, Phospho-SAPK/JNK Rabbit mAb and Phospho-Erk1/2 Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from Jurkat, C6, NIH/3T3 and COS cells, untreated or treated as indicated, using Phospho-p38 MAPK (Thr180/Tyr182) (3D7) Rabbit mAb (upper) or p38 MAPK Antibody #9212 (lower).

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells, untreated (blue) or anisomycin-treated (green), using Phospho-p38 MAPK (Thr180/Tyr182) (3D7) Rabbit mAb compared to a nonspecific negative control antibody (red).


IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells, untreated (left) or anisomycin-treated (right), using Phospho-p38 MAPK (Thr180/Tyr182)(3D7) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).

Background

p38 MAP kinase (MAPK), also called RK (1) or CSBP (2), is the mammalian orthologue of the yeast HOG kinase that participates in a signaling cascade controlling cellular responses to cytokines and stress (1-4). Four isoforms of p38 MAPK, p38α, β, γ (also known as Erk6 or SAPK3), and δ (also known as SAPK4) have been identified. Similar to the SAPK/JNK pathway, p38 MAPK is activated by a variety of cellular stresses including osmotic shock, inflammatory cytokines, lipopolysaccharide (LPS), UV light, and growth factors (1-5). MKK3, MKK6, and SEK activate p38 MAPK by phosphorylation at Thr180 and Tyr182. Activated p38 MAPK has been shown to phosphorylate and activate MAPKAP kinase 2 (3) and to phosphorylate the transcription factors ATF-2 (5), Max (6), and MEF2 (5-8).SB203580 (4-(4-fluorophenyl)-2-(4-methylsulfinylphenyl)-5-(4-pyridyl)-imidazole) is a selective inhibitor of p38 MAPK. This compound inhibits the activation of MAPKAPK-2 by p38 MAPK and subsequent phosphorylation of HSP27 (9). SB203580 inhibits p38 MAPK catalytic activity by binding to the ATP-binding pocket, but does not inhibit phosphorylation of p38 MAPK by upstream kinases (10).

  1. Rouse, J. et al. (1994) Cell 78, 1027-1037.
  2. Han, J. et al. (1994) Science 265, 808-811.
  3. Lee, J.C. et al. (1994) Nature 372, 739-746.
  4. Freshney, N.W. et al. (1994) Cell 78, 1039-1049.
  5. Raingeaud, J. et al. (1995) J. Biol. Chem. 270, 7420-7426.
  6. Zervos, A.S. et al. (1995) Proc. Natl. Acad. Sci. USA 92, 10531-10534.
  7. Zhao, M. et al. (1999) Mol. Cell. Biol. 19, 21-30.
  8. Yang, S.H. et al. (1999) Mol. Cell. Biol. 19, 4028-4038.
  9. Cuenda, A. et al. (1995) FEBS Lett 364, 229-33.
  10. Kumar, S. et al. (1999) Biochem Biophys Res Commun 263, 825-31.

Application References

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Companion Products

Rabbit Monoclonals Produced Using Epitomics® Technology, U.S. Patent No. 5,675,063.

For Research Use Only. Not For Use In Diagnostic Procedures.

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