Product Pathways - MAPK Signaling

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p38 MAPK Antibody #9212

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PhosphoSitePlus^® protein, site, and accession data: p38-alpha, p38-beta, p38-gamma

Applications	Reactivity	Sensitivity	MW (kDa)	Source
W IHC-P IF-IC F	H M R Mk GP (C)	Endogenous	43	Rabbit

Applications Key: W=Western Blotting IHC-P=Immunohistochemistry (Paraffin) IF-IC=Immunofluorescence (Immunocytochemistry) F=Flow Cytometry
Reactivity Key: H=Human M=Mouse R=Rat Mk=Monkey C=Chicken GP=Guinea Pig
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

9212:: Flow, IHC / Paraffin, Immunofluorescence, Western Blotting

Specificity / Sensitivity

p38 MAP Kinase Antibody detects endogenous levels of total p38α, -β or -γ MAPK protein. This antibody does not recognize p38δ, JNK/SAPK or p44/42 MAPK.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the sequence of human p38 MAPK. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence^® Control siRNA (Unconjugated) #6568 (-), SignalSilence^® p38 MAPK siRNA I #6564 (+) or SignalSilence^®p38 MAPK siRNA II (+), using p38 MAPK Antibody #9212 and α-Tubulin (11H10) Rabbit mAb #2125. The p38 MAPK antibody confirms silencing of p38 MAPK expression while the α-Tubulin (11H10) Rabbit mAb is used to control for loading and specificity of p38 MAPK siRNA.

Western Blotting

Western blot analysis of extracts from C6 cells, untreated or anisomycin-treated, and NIH/3T3 cells, untreated or UV-treated, using phospho-p38 MAPK (Thr180/Tyr182) Antibody #9211 (upper) or p38 MAPK Antibody (lower).

Western Blotting

Western blot analysis of extracts from NIH/3T3 cells, untreated or UV-treated for the indicated times, using Phospho-p38 MAPK (Thr180/Tyr182) Antibody #9211 (upper) or p38 MAPK Antibody (lower).

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, showing nuclear and cytoplasmic localization, using p38 MAPK Antibody.

Flow Cytometry

Flow cytometric analysis of Jurkat cells, using p38 MAPK Antibody (blue) compared to a nonspecific negative control antibody (red).

IF-IC

Confocal immunofluorescent images of HeLa cells-/+ UV light labeled with #9212 p38 MAPK (green) exhibiting cytoplasmic localization 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).

Application References

Miyaji, M. et al. (2009) Proc Natl Acad Sci U S A 106, 14502-7. Applications: Western Blotting
Hsu, J.T. et al. (2007) Am J Physiol Heart Circ Physiol 292, H2982-7. Applications: Western Blotting
Kojima, R. et al. (2004) Biochem J 380, 783-94. Applications: Western Blotting
Matsumoto, M. et al. (2004) J Biol Chem 279, 45969-79. Applications: IF-IC (In Cells)
Matsumoto, M. et al. (2000) Involvement of p38 mitogen-activated protein kinase signaling pathway in osteoclastogenesis mediated by receptor activator of NF-kappa B ligand (RANKL). J. Biol. Chem. 275, 31155-31161. Applications: Western Blotting
Chen, C. H. et al. (2000) Cyclic AMP activates p38 mitogen-activated protein kinase in Th2 cells: phosphorylation of GATA-3 and stimulation of Th2 cytokine gene expression. J. Immunol. 165, 5597-5605. Applications: Western Blotting
Mudgett, J. S. et al. (2000) Essential role for p38alpha mitogen-activated protein kinase in placental angiogenesis. Proc. Natl. Acad. Sci. USA 97, 10454-10459. Applications: IHC-P (paraffin) Western Blotting
Rossi, A. et al. (2000) Anti-inflammatory cyclopentenone prostaglandins are direct inhibitors of IkappaB kinase. Nature 403, 103-108. Applications: Western Blotting
Zhuang, S. et al. (2000) p38 mitogen-activated protein kinase mediates bid cleavage, mitochondrial dysfunction, and caspase-3 activation during apoptosis induced by singlet oxygen but not by hydrogen peroxide. J. Biol. Chem. 275, 25939-25948. Applications: Western Blotting
Rohani, M.G. et al. (2010) BMC Immunol 11, 53. Applications: Western Blotting
Gehani, S.S. et al. (2010) Mol Cell 39, 886-900. Applications: Western Blotting
Lee, W.H. et al. (2011) J Biol Chem 286, 21577-87. Applications: Western Blotting

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