Product Pathways - MAPK Signaling

Phospho-p38 MAPK Pathway Sampler Kit #9913

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Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IHC-F=Immunohistochemistry (Frozen)  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  Dr=Drosophila  Z=Zebra Fish  Dg=Dog

Specificity / Sensitivity

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

Western Blotting

Western blot analysis of extracts from HeLa cells, using Phospho-HSP27 (Ser82) Antibody #2401 (upper) and control HSP27 antibody (lower). HeLa cells were incubated at 42°C for 0-2 hours as indicated.

Western Blotting

Western blot analysis of extracts from HeLa cells stimulated with anisomycin for indicated times, using Phospho-MAPKAPK-2 (Thr334) Antibody #3041 (upper) and control MAPKAPK-2 antibody (lower).

Western Blotting

Western blot analysis of Elk-1 fusion protein expressed from E. coli with or without phosphorylation by purified Erk2 enzyme, using Phospho-Elk-1 (Ser383) Antibody #9181 (upper) or control Elk-1 Antibody #9182 (lower).

Western Blotting

Western blot analysis of extracts from UV treated NIH/3T3 cells, using Phospho-p38 MAP Kinase (Thr180/Tyr182) Antibody #9211 (upper) or control p38 MAP Kinase Antibody #9212 (lower).

Western Blotting

Western blot analysis of C6 cell extracts treated or untreated with anisomycin, using Phospho-p38 MAP Kinase (Thr180/Tyr182) Antibody #9211 (upper) and p38 MAP Kinase Antibody #9212 (lower).

Western Blotting

Western blot analysis of extracts from anisomycin treated NIH/3T3 cells, using Phospho-ATF-2 (Thr71) Antibody #9221 (upper) or control ATF-2 Antibody #9222 (lower).

Western Blotting

Western blot analysis of anisomycin treated NIH/3T3 cell extracts, using Phospho-MKK3/MKK6 (Ser189/207) Antibody #9231 (upper) and control MKK3 Antibody #9232 (lower).

IHC-P (paraffin)

Immunohistochemical staining of phosphorylated HSP27 in paraffin-embedded human tonsil showing cytoplasmic localization, using Phospho-HSP27 (Ser82) Antibody #2401.

IHC-P (paraffin)

Immunohistochemical staining of paraffin-embedded human breast carcinoma showing nuclear localization of phosphorylated MAPKAPK-2, using Phospho-MAPKAPK-2 (Thr334) Antibody #3041.

IHC-P (paraffin)

Immunohistochemical staining of paraffin-embedded human breast carcinoma, using Phospho-ATF-2 (Thr71) Antibody #9221 showing nuclear localization of phosphorylated ATF-2.

IHC-FL (floating)

Confocal images of double immunostaining using Phospho-ATF-2 (Thr71) Antibody #9221 (green) and calbindin antibody (red) in the hippocampus, using the sections from control and 24 hours of reperfusion following 15 minute transient cerebral ischemia in rats. Yellow represents overlay of red and green. (Provided by Dr. Bingren Hu, University of Miami School of Medicine, Florida.)

IHC-FL (floating)

Confocal images of double immunostaining using Phospho-ATF-2 (Thr71) Antibody #9221 (green) and calbindin (red) in the hippocampal CA1, using brain sections from control and 24 hour of reperfusion following 15 minute transient cerebral ischemia in rats. Yellow represents overlay of red and green. (Provided by Dr. Bingren Hu, University of Miami School of Medicine, Florida.)

Source / Purification

Polyclonal antibodies are produced by immunizing rabbits with synthetic phospho-peptides corresponding to residues surrounding Ser383 of human Elk-1, Thr180/Tyr182 of human p38 MAPK, Ser189/207 of human MKK3, Thr71 of human ATF-2, Ser82 of human HSP27 or Thr334 of human MAPKAPK-2. Antibodies are purified by protein A and peptide affinity chromatography.

Background

p38 MAP kinase (MAPK), also called RK (1) or CSBP (2), is the mammalian orthologue of the yeast HOG kinase which participates in a signaling cascade controlling cellular responses to cytokines and stress (1-4). Four isoforms of p38 MAP kinase, 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 MAP kinase is activated by a variety of cellular stresses including osmotic shock, inflammatory cytokines, lipopolysaccharides (LPS), UV light and growth factors (1-5). MKK3, MKK6 and SEK activate p38 MAP kinase by phosphorylation at Thr180 and Tyr182. Activated p38 MAP kinase 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).

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

Application References

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