CST spho-p38 MAPK （Thr180/Tyr182） （28B10） Mouse mAb

Applications Key: W=Western Blotting IP=Immunoprecipitation IF-IC=Immunofluorescence （Immunocytochemistry） F=Flow Cytometry

Reactivity Key: H=Human M=Mouse R=Rat Mk=Monkey Z=Zebrafish 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

9216:

Flow, Immunofluorescence, Immunoprecipitation, Western Blotting

Specificity / Sensitivity

Phospho-p38 MAPK （Thr180/Tyr182） （28B10） Mouse mAb detects p38 MAPK only when activated by dual phosphorylation at Thr180 and Tyr182. This antibody does not significantly cross-react with the corresponding phosphorylated forms of either p44/42 MAPK （Erk1/2） or SAPK/JNK. It does not detect nonphosphorylated p38 MAPK.

Source / Purification

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

Western Blotting

Western blot analysis of extracts from HeLa and NIH/3T3 cells, untreated or treated as indicated, using Phospho-p38 MAPK （Thr180/Tyr182） （28B10） Mouse mAb （upper） or p38 MAPK Antibody #9212 （lower）.

Flow Cytometry

Flow cytometric analysis of Jurkat cells, untreated （green） or anisomycin-treated （blue）, using Phospho-p38 MAPK （Thr180/Tyr182） （28B10） Mouse mAb compared to a nonspecific negative control antibody （red）.

IF-IC

Confocal immunofluorescent analysis of HeLa cells either untreated （left） or anisomycin treated （right） labeled with Phospho-p38 MAP Kinase （Thr180/Tyr182） （28B10） Mouse mAb （green）. Actin filaments have been labeled with Alexa Fluor Phalloidin 555 （red）.

CST spho-p38 MAPK （Thr180/Tyr182） （28B10） Mouse mAb

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）.

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

Application ReferencesGehani, S.S. et al. （2010） Mol Cell 39, 886-900. Applications: Western Blotting Xu, P. and Derynck, R. （2010） Mol Cell 37, 551-66. Applications: Western Blotting Wu, M. et al. （2009） Diabetes Metab Res Rev 25, 279-86. Applications: Western Blotting Basak, C. et al. （2005） J Biol Chem 280, 4279-88. Applications: Western Blotting

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

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