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Technical Data Sheet


p38 MAPK Phospho-Regulation
Immunocytochemistry Kit


Kit Components
Applications: WB = Western blot, E = ELISA, ICC = Immunocytochemistry, IP = Immunoprecipitation, IHC = Immunohistochemistry Species: H = Human, R = Rat, M = Mouse, C = Chicken, F = Fish

Product References
PP3411 Alam, M.S. et al. (2015) Nat Medicine 21:1337. WB, IF: human T-cells, CD3 activated
PP3411 Lanna, A. et al. (2014) Nat Immunol. 15(10):965. WB: human T-cells, CD3 activated
PM1391 Li, W. et al. (2009) AJP Cell Phys. 297:C706. WB: mouse C2C12 cells
PM1391 Chambers, M.A. et al. (2009) J Physiol. 587:3363. WB: mouse muscle, C2C12
Kit Summary
The p38 MAPK phospho-regulation kit can be used for immunocytochemical co-localization of p38 MAPK phosphorylation at Thr-180/Tyr-182 and Tyr-323 relative to the total expression of p38 MAPK. The kit includes rabbit polyclonal and mouse monoclonal antibodies along with Goat-anti-Rabbit conjugated to DyLight® 594 and Goat anti-Mouse conjugated to DyLight® 488 for dual labeling experiments.

Immunocytochemical labeling of p38 MAPK in control and pervanadate-treated mouse C2C12 cells. The cells were labeled with mouse monoclonal p38α MAPK (PM1381), p38 MAPK (Thr-180/Tyr-182) (PM1391), and rabbit polyclonal p38 MAPK (Tyr-323) (PP3411). The antibodies were detected using appropriate secondary conjugated to DyLight® 488 or DyLight® 594.

p38 MAP kinase (MAPK), also called RK, CSBP, and SAPK2a, is the mammalian orthologue of the yeast HOG kinase. This family of kinases participates in signaling cascades that control cellular responses to cytokines and stress. Four isoforms of p38 MAPK (α,β,γ,δ) have been identified. Similar to the SAPK/JNK pathway, p38 MAPK is activated by a variety of cellular stresses including osmotic shock, inflammatory cytokines, lipopolysaccharides, UV light, and growth factors. MKK3 and SEK activate p38 MAPK by dual phosphorylation at Thr-180/Tyr-182. Activated p38 MAPK has been shown to phosphorylate and activate MAPKAP kinase 2 and to phosphorylate the transcription factors ATF-2, Max, and MEF2. T cells possess an alternate pathway for p38 activation where stimulation of the antigen receptor (TCR) induces phosphorylation of p38 on Tyr-323. This site is required for TCR-mediated phosphorylation of Thr-180 and catalytic activity. Thus, Tyr-323 may also have important roles in regulating p38 MAP kinase pathways.

Background References
Han, J. et al. (1994) Science 265:808.
Rouse, J. et al. (1994) Cell 78:1027.
Lee, J. C. et al. (1994) Nature 372:739.
Freshney, N. W. et al. (1994) Cell 78:1039.
Raingeaud, J. et al. (1995) J. Biol. Chem. 270:7420.
Salvador, J.M. et al. (2005) Nat Immunol. 6(4):390.
Jirmanova, L. et al. (2009) Blood 113(10):2229.
Buffer and Storage
Mouse monoclonal, rabbit polyclonal, and secondary reagents are supplied in phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. Store at –20°C. Stable for 1 year.
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