Cell Signaling Technology

Product Pathways - MAPK Signaling

Phospho-p38 MAPK (Thr180/Tyr182) (12F8) Rabbit mAb #4631

Applications Reactivity Sensitivity MW (kDa) Isotype
W IHC-P IF-IC H M R Mk Dm (Hm) (Mi) (Z) Endogenous 43 Rabbit IgG

Applications Key:  W=Western Blotting  IHC-P=Immunohistochemistry (Paraffin)  IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:  H=Human  M=Mouse  R=Rat  Hm=Hamster  Mk=Monkey  Mi=Mink  Dm=D. melanogaster  Z=Zebrafish
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) (12F8) Rabbit mAb detects endogenous levels of p38 MAPK only when dually phosphorylated at threonine 180 and tyrosine182. It will also react with p38 singly phosphorlyated at Thr180. 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

Western blot analysis of extracts from Jurkat, NIH/3T3 and PC12 cells, untreated or treated as indicated, using Phospho-p38 MAPK (Thr180/Tyr182) (12F8) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemistry of paraffin-embedded lung carcinoma, showing nuclear localization, using Phospho-p38 MAPK (Thr180/Tyr182) (12F8) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemistry of paraffin-embedded colon carcinoma, showing nuclear localization, using Phospho-p38 MAPK (Thr180/Tyr182) (12F8) Rabbit mAb.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemistry of paraffin-embedded human glioblastoma, untreated (left) or CIP phosphatase-treated (right), using Phospho-p38 MAPK (Thr180/Tyr182) (12F8) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemistry of paraffin-embedded breast carcinoma, using Phospho-p38 MAPK (Thr180/Tyr182) (12F8) Rabbit mAb (left) or the same antibody preincubated with antigen phospho-peptide (right).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemistry of paraffin-embedded NIH/3T3 cells, untreated (left) or anisomycin-treated (right), using Phospho-p38 MAPK (Thr180/Tyr182) (12F8) Rabbit mAb.


IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells -/+ UV light, labeled with Phospho-p38 MAPK (green). Absence of staining in untreated cells (left) and nuclear localization in treated cells (right). Red = Actin filaments (phalloidin).

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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