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Technical Data Sheet

MK6050

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MAP Kinase Activation
Antibody Sampler Kit
Price

$395

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
EM2331 Kyjacova, L. et al. (2015) Cell Death Differ. 22(6):898. WB: human DU145
EM2061 Park, K. et al. (2013) Mol Cell Biol. 33(4):752. WB: human keratinocytes
PM1391 Chambers, M.A. et al. (2009) J Physiol. 587:3363. WB: mouse muscle, C2C12
PM1391 Li, W. et al. (2009) AJP Cell Phys. 297:C706. WB: mouse C2C12 cells
Kit Summary
The MAP kinase activation antibody sampler kit can be used to examine the activation of three major MAPKs: p38, ERK, and JNK. The kit also includes antibodies to monitor total expression levels of p38, ERK1/2, and JNK1.
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Western blot analysis of A431 cells serum starved overnight (lanes 1 & 3) then treated with pervanadate (1 mM) for 30 min. (lanes 2 & 4). The blot was probed with anti-p38α (lanes 1 & 2) or anti-p38 (T180/Y182) (lanes 3 & 4).

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Western blot analysis of human A431 cells untreated (lanes 1 & 4), treated with 100 nM calyculin A for 30 min. (lanes 2 & 5), or 100 ng/ml EGF for 60 min. (lanes 3 & 6). The blots were probed with anti-ERK1 (lanes 1, 2, & 3) or anti-ERK1/2 (Thr-202/Tyr-204) (lanes 4, 5, & 6).

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Background
Mitogen-activated protein kinases (MAPKs) are a widely conserved family of serine/threonine protein kinases that include three major MAPKs: p38, JNK, and ERK. These MAPKs are involved in many cellular programs such as cell proliferation, differentiation, motility, and death. Upon stimulation, the MAPKs are activated through a sequential protein kinase cascade consisting of MAPKKKs activating MAPKKs followed by dual phosphorylation and activation of MAPKs. p38 and JNK MAPKs participate in signaling cascades that control cellular responses to cytokines and stress. Activated p38 MAPK has been shown to phosphorylate and activate MAPKAP kinase 2 and to phosphorylate the transcription factors ATF-2, Max, and MEF2. Active JNK dimers can translocate to the nucleus to regulate transcription through phosphorylation of c-Jun, ATF-2, and other transcription factors. The ERK1/2 (p44/42) signaling pathway can be activated in response to a diverse range of extracellular stimuli including mitogens, growth factors, and cytokines. Activated ERKs have several downstream targets including p90RSK and Elk-1.

Background References
Freshney, N. W. et al. (1994) Cell 78:1039.
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.
Raingeaud, J. et al. (1995) J. Biol. Chem. 270:7420.
Whitmarsh, A.J. & Davis, R.J. (1998) Trends Biochem. Sci. 23:481.
Kyriakis, J.M. (1999) J. Biol. Chem. 274:5259.
Davis, R.J. (1999) Biochem. Soc. Symp. 64:1.
Ichijo, H. (1999) Oncogene 18:6087.
Roux, P.P. & Blenis, J. (2004) Microbiol Mol Biol Rev 68:320.
Murphy, L.O. & Blenis, J. (2006) Trends Biochem Sci 31:268.
Owens, D.M. & Keyse, S.M. (2007) Oncogene 26:3203.
Buffer and Storage
Mouse monoclonal 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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