Catalog # EM2061

ERK1 (Thr-202/Tyr-204)[conserved], phospho-specific Antibody

Mouse Monoclonal

Application / Dilution

ELISA	1:2000
ICC	1:100
WB	1:1000

Size 100 μl

Species Reactivity Hu, Rt, Ms

MW 44 kDa

Isotype IgG1

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.

References

Owens, D.M. & Keyse, S.M. (2007) Oncogene 26:3203.
Murphy, L.O. & Blenis, J. (2006) Trends Biochem Sci 31:268.
Roux, P.P. & Blenis, J. (2004) Microbiol Mol Biol Rev 68:320.

Western blot analysis of human A431 epithelial cells untreated (lanes 1 & 4) or 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 (C-terminal region) (lanes 1, 2, & 3) or anti-ERK1/2 (Thr-202/Tyr-204) (lanes 4, 5, & 6).

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).

Clone M206 was generated from a dual phosphorylated ERK1 (Thr-202/Tyr-204) synthetic peptide (coupled to carrier protein) corresponding to amino acids surrounding Thr-202 and Tyr-204 in human ERK1. This sequence is conserved in rat and mouse ERK1, and is highly conserved in ERK2.

*For more information, see UniProt Accession P27361

Mouse monoclonal antibody purified with protein A chromatography 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.

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.

The antibody detects 42 and 44 kDa* proteins corresponding to ERK1 and ERK2 on SDS-PAGE immunoblots of human A431 epithelial cells stimulated with EGF or calyculin A. It does not detect bands in control cells.

*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:

Elizondo, DM et al. (2019) Front Immunol. 10:173. (WB: mouse dendritic cells)
Park, K. et al. (2013) Mol Cell Biol. 33(4):752. (WB: human keratinocytes)

This kit contains: