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Technical Data Sheet

EK7620

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ERK1/2 Phospho-Regulation
Immunocytochemistry Kit
Price

$355

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
Kit Summary
The ERK1/2 phospho-regulation kit may be used to examine the pattern of phosphorylation of ERK1 at the dual site (Thr-202/Tyr-204) relative to the total ERK1 pattern. The kit includes monoclonal antibodies to ERK1 (C-terminal region) and ERK1/2 (Thr-202/Tyr-204), as well as Goat anti-mouse secondary reagents conjugated to DyLight® 488 or DyLight® 594 for detection using green (493Ex/518Em) or red (593Ex/618Em) filter sets.
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Immunocytochemical labeling of phosphorylated ERK1 in paraformaldehyde-fixed and NP-40-permeabilized rat A7r5 cells treated with calyculin A. The fixed cells were labeled with mouse monoclonal antibodies to anti-ERK1 (EM2331) and anti-ERK1/2 (Thr-202/Tyr-204) (EM2061). The antibodies were detected using Goat anti-Mouse secondary antibodies conjugated to DyLight® 488 (left) and DyLight® 594 (right).

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Background
Mitogen-activated protein kinases (MAPKs) are a widely conserved family of serine/threonine protein kinases involved in many cellular programs such as cell proliferation, differentiation, motility, and death. 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. Upon stimulation, a sequential three-part protein kinase cascade is initiated, consisting of a MAP kinase kinase kinase (MAPKKK), a MAP kinase kinase (MAPKK), and a MAP kinase (MAPK). Multiple ERK1/2 MAPKKKs have been identified, including members of the Raf family as well as Mos and Tpl2/Cot. MEK1 and MEK2 are the primary MAPKKs in this pathway. MEK1 and MEK2 activate ERK1 and ERK2 through phosphorylation of activation loop residues Thr-202/Tyr-204 and Thr-185/Tyr-187, respectively. ERK1/2 are negatively regulated by a family of dual-specificity (Thr/Tyr) MAPK phosphatases. Several downstream targets of ERK1/2 have been identified, including p90RSK and the transcription factor Elk-1.

Background References
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 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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