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Technical Data Sheet

EP4101

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ERK1(T207)/ERK2(T188)[conserved], phospho-specific
Rabbit Polyclonal
Price
Size
Species Reactivity
MW

$295
100 μl
Hu, Rt, Ms, Ck, F
42 kDa

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Western blot analysis of human A431 epithelial cells treated with 100 nM calyculin A for 30 min. (lanes 1, 3, 5, & 7) then the blot was treated with lambda phosphatase (lanes 2, 4, 6, & 8). The blots were probed with polyclonal anti-ERK2 (a.a. 181-195) (lanes 1 & 2), anti-ERK2 (Thr-188) (lanes 3 & 4), anti-ERK1/2 (Thr-202/Tyr-204) (lanes 5 & 6), or monoclonal anti-ERK1 (C-terminal region) (lanes 7 & 8).

Application
Dilution


ELISA
1:2000


WB
1:1000



End user should determine optimal dilution for their particular applications and experiments.Western blot membranes were incubated with diluted antibody in 5% non-fat milk, PBS, 0.04% Tween20 for 1hour at room temperature.
Product References
H. Huang, et al. (2015) Cardiovasc Res. 108(1):50. WB: mouse heart
Lu, J. et al. (2013) Basic Res Cardiol. 108(2):326. WB fluorescence: mouse heart
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Background
The ERK1/2 (p44/42) MAPK 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 MAP kinase cascade is initiated, consisting of a MAP kinase kinase kinase (MAPKKK), a MAP kinase kinase (MAPKK), and a MAP kinase (MAPK). Activation of the MAPKs, ERK1 and ERK2, leads to phosphorylation of activation loop residues Thr-202/Tyr-204 and Thr-185/Tyr-187, respectively. In addition to dual phosphorylation, ERK1 and 2 are autophosphorylated on Thr-207 or Thr-188, respectively. This phosphorylation is required for nuclear translocation of ERK, and leads to phosphorylation of several nuclear proteins involved in cardiac hypertrophy. Mouse models with mutation of Thr-188 in ERK2 show that this site is critical for ERK-mediated cardiac hypertrophy. Thus, phosphorylation of Thr-188 in ERK2 may be important for controlling the nuclear functions of activated ERK1 and ERK2.


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.
Lorenz, K. et al. (2009) Nat Med. 15(1):75.
Immunogen
Phospho-ERK2 (Thr-188) synthetic peptide (coupled to carrier protein) corresponds to amino acids surrounding Thr-188 in mouse ERK2. This sequence is conserved in human, rat, chicken, and fish ERK2, and is highly conserved in ERK1 (Thr-207), ERK5 (Thr-224), and ERK7 (Thr-180).
Buffer and Storage
Rabbit polyclonal, affinity-purified antibody is supplied in 100µl phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. Store at –20°C. Stable for 1 year.
Specificity
The antibody detects 42 and 44 kDa* proteins corresponding to ERK1 (Thr-207) and ERK2 (Thr-188) on SDS-PAGE immunoblots of human A431 epithelial cells stimulated with calyculin A. It does not detect these ERK proteins in control cells or in blots treated with lambda phosphatase.

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