Cell Signaling Technology

Product Pathways - MAPK Signaling

Phospho-p38 MAPK Pathway Sampler Kit #9913

Kit Includes Quantity Applications Reactivity MW (kDa) Isotype
Phospho-p38 MAPK (Thr180/Tyr182) (D3F9) XP® Rabbit mAb #4511 40 µl W IP IHC-P IF-IC F H M R Mk Mi Pg Sc (Hm) (C) (Z) (B) 43 Rabbit IgG
Phospho-MSK1 (Thr581) Antibody #9595 40 µl W IP IHC-P H M 90 Rabbit
Phospho-MKK3 (Ser189)/MKK6 (Ser207) (22A8) Rabbit mAb #9236 40 µl W IP H M R Mk 40 P-MKK3, 41 P-MKK6 Rabbit IgG
Phospho-ATF-2 (Thr71) (11G2) Rabbit mAb #5112 40 µl W F H M R Mk 70 Rabbit IgG
Phospho-MAPKAPK-2 (Thr334) (27B7) Rabbit mAb #3007 40 µl W IHC-P IF-IC F H M R Mk 49 Rabbit IgG
Phospho-HSP27 (Ser82) (D1H2) XP® Rabbit mAb #9709 40 µl W IHC-P IF-IC F H M (R) (Hm) (B) (Dg) (Hr) 27 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl Goat

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  Hm=Hamster  Mk=Monkey  C=Chicken  Mi=Mink  Z=Zebrafish  B=Bovine  Dg=Dog  Pg=Pig  Sc=S. cerevisiae  Hr=Horse
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Specificity / Sensitivity

Each antibody in the Phospho-p38 MAPK Pathway Sampler Kit recognizes only the phosphorylated form of its specific target.

Western Blotting

Western Blotting

Western blot analysis of extracts from untreated or UV+TPA-treated HeLa and COS cells, using Phospho-MAPKAPK-2 (Thr334) (27B7) Rabbit mAb #3007 (upper), or MAPKAPK-2 Antibody #3042 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from COS and 293 cells, untreated or UV-treated, using Phospho-p38 MAPK (Thr180/Tyr182) (D3F9) Rabbit mAb #4511 (upper) or p38 MAPK Antibody #9212 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from untreated or UV-treated COS cells, NIH/3T3 cells and C6 cells, using Phospho-ATF-2 (Thr71) (11G2) Rabbit mAb #5112 (upper), or ATF-2 (20F1) Rabbit mAb #9226 (lower).


Western Blotting

Western Blotting

Western blot analysis of extracts from untreated or UV-treated COS, NIH/3T3 and C6 cells, using Phospho-MKK3/6 (Ser189/207) (22A8) Rabbit mAb #9236.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293 cells untreated or treated with UV and HeLa cells untreated or treated with calyculin A, using Phospho-MSK1 (Thr581) Antibody #9595.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa or HT-29 cells, untreated (-) or treated (+) with either UV (40 mJ/cm2 with 30 min recovery) or anisomycin (25 μg/mL, 30 min), using Phospho-HSP27 (Ser82) (D1H2) XP® Rabbit mAb #9709.


Description

The Phospho-p38 MAPK Pathway Sampler Kit provides an economical means to evaluate the activation status of multiple members of the p38 MAPK pathway, including phosphorylated MSK1, p38 MAPK, MKK3/MKK6, ATF-2, HSP27 and MAPKAPK-2. The kit includes enough primary and secondary antibodies to perform four Western blot experiments.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with synthetic phosphopeptides corresponding to residues surrounding Thr581 of human MSK1, Thr180/Tyr182 of human p38 MAPK, Ser189 of human MKK3, Thr71 of human ATF-2, Ser82 of human HSP27 or Thr334 of human MAPKAPK-2.

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).

Four residues (Thr25, Thr222, Ser272 and Thr334) of MAPKAPK-2 are phosphorylated by p38 in an in vitro kinase assay (3). Phosphorylation at Thr222, Ser272 and Thr334 seems to be essential for the activity of MAPKAPK-2 (3). Activated MAPKAPK-2 can in return phosphorylate HSP27 at serines 15, 78 and 82 (3,9). Phosphorylation of HSP27 causes a change in the tertiary structure of HSP27, which shifts from large homotypic multimers to dimmers and monomers (10). It has been illustrated that phosphorylation and increased concentration of HSP27 modulate actin polymerization and reorganization (11,12).

  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.
  11. Landry, . et al. (1992) J. Biol. Chem. 267, 794-803.
  12. Rogalla, . et al. (1999) J. Biol. Chem. 274, 18947-18956.
  13. Lavoie, J. et al. (1993) J. Biol. Chem. 268, 24210-24214.
  14. Rousseau, S. et al. (1997) Oncogene 15, 2169-2177.

Application References

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Protocols

Companion Products

Selected rabbit monoclonal antibodies are produced under license (granting certain rights including those under U. S. Patent No. 5,675,063 and/or U.S.S.N. 11/476,277) from Epitomics, Inc.

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

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