Catalog # EK7620

ERK1/2 Phospho-Regulation Staining Kit

Immunocytochemistry Kit



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.

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Murphy, L.O. & Blenis, J. (2006) Trends Biochem Sci 31:268.
Owens, D.M. & Keyse, S.M. (2007) Oncogene 26:3203.

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).

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.

The products are are safely shipped at ambient temperature for both domestic and international shipments. Each product is guaranteed to match the specifications as indicated on the corresponding technical data sheet. Please store at -20C upon arrival for long term storage.

*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot mobilities of known proteins with similar MW.

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

This kit contains:

ERK1 (C-terminal region) Mouse mAb50 μlWB, E, ICC, IHCHu, Rt, Ms1:100
ERK1 (Thr-220)/ (Tyr-204)[conserved], phospho-specific Mouse mAb50 μlWB, E, ICCHu, Rt, Ms 1:100
Anti-Mouse Ig:DyLight® 488 Goat pAb100 μlICC, IHCMs1:200
Anti-Mouse Ig:DyLight® 594 Goat pAb100 μlICC, IHCMs1:200

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.