Catalog # EK6440

ERK1/2 Phospho-Regulation Antibody Kit

Antibody Sampler Kit

DATASHEET  




$395

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 ERK.

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.

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).

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 rabbit polyclonal antibodies are supplied in phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. The secondary reagents are supplied in the same buffer without azide. Store all 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
EP4101 H. Huang, et al. (2015) Cardiovasc Res. 108(1):50. WB: mouse heart
EM2061 Park, K. et al. (2013) Mol Cell Biol. 33(4):752. WB: human keratinocytes
EP4101 Lu, J. et al. (2013) Basic Res Cardiol. 108(2):326. WB fluorescence: mouse heart
RS3251 Kawasaki, H. et al. (2013) World J Gastroenter. 19(17):2629. WB, ICC: mouse intestinal myofibroblasts
MS3001 Estrada-Bernal, A. et al. (2011) J Neurooncol. 102:353. Western Blot
RS3251 Estrada-Bernal, A. et al. (2011) J Neurooncol. 102:353. Western Blot

This kit contains:

CATALOG#DESCRIPTIONSIZEAPPLICATIONSSPECIES REACTIVITYMW (kDa)
ERK1 (C-terminal region) Mouse mAb50 μlWB, E, ICC, IHCHu, Rt, Ms44
ERK1 (Thr-202/Tyr-204)[conserved] Mouse mAb50 μlWB, E, ICCHu, Rt, Ms44
ERK2 (a.a. 181-195)[conserved site] Rabbit pAb 50 μlWB, EHu, Rt, Ms, Ck, F42
ERK1 (Thr-207)/ERK2 (Thr-188)[conserved], phospho-specific Rabbit pAb50 μlWB, EHu, Rt, Ms, Ck, F42
Anti-Mouse Ig:HRP Donkey pAb 100 μlWB, EMs
Anti-Rabbit Ig Light ChainSpecific:HRP Mouse mAb 100 μlWB, E, ICC, IHCRb
KIT SUMMARY

The ERK phospho-regulation antibody sampler kit can be used to examine ERK1 phosphorylation at Thr-202/Tyr-204 and Thr-207. These phosphorylation sites are also conserved in ERK2. A mouse monoclonal antibody to ERK1 and a rabbit polyclonal antibody to ERK2 are also included for examining total ERK expression levels.