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 Pho 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）.

 

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

• Gehani, 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

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Companion Products

• 9913 Phospho-p38 MAPK Pathway Sampler Kit
• 9213 p38 MAPK Control Cell Extracts
• 4511 Phospho-p38 MAPK （Thr180/Tyr182） （D3F9） XP^® Rabbit mAb
• 9215 Phospho-p38 MAPK （Thr180/Tyr182） （3D7） Rabbit mAb
• 2371 p38α MAPK （7D6） Rabbit mAb
• 9217 p38α MAPK （5F11） Mouse mAb
• 9228 p38α MAPK （L53F8） Mouse mAb
• 2339 p38β MAPK （C28C2） Rabbit mAb
• 2308 p38δ MAPK （10A8） Rabbit mAb
• 4631 Phospho-p38 MAPK （Thr180/Tyr182） （12F8） Rabbit mAb
• 2307 p38γ MAPK Antibody
• 9211 Phospho-p38 MAPK （Thr180/Tyr182） Antibody
• 9212 p38 MAPK Antibody
• 9218 p38α MAPK Antibody
• 9214 p38δ MAPK Antibody
• 9210 PhosphoPlus^® p38 MAPK （Thr180/Tyr182） Antibody Kit
• 9219 Phospho-p38 MAPK （Thr180/Tyr182） Mouse mAb （Sepharose Bead Conjugate）
• 7072 Phototope^®-HRP Western Blot Detection System, Anti-mouse IgG, HRP-linked Antibody
• 7076 Anti-mouse IgG, HRP-linked Antibody
• 7720 Prestained Protein Marker, Broad Range （Premixed Format）
• 7727 Biotinylated Protein Ladder Detection Pack
• 7003 20X LumiGLO^® Reagent and 20X Peroxide

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

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