Product Pathways - MAPK Signaling
Phospho-MEK1 (Thr286) Antibody #9127
Applications | Reactivity | Sensitivity | MW (kDa) | Source |
---|---|---|---|---|
W IP IF-IC F | H R Mk (M) | Endogenous | 45 | Rabbit |
Applications Key:
W=Western Blotting
IP=Immunoprecipitation
IF-IC=Immunofluorescence (Immunocytochemistry)
F=Flow Cytometry
Reactivity Key:
H=Human
M=Mouse
R=Rat
Mk=Monkey
Species cross-reactivity is determined by Western blot.
Protocols
Specificity / Sensitivity
Phospho-MEK1 (Thr286) Antibody detects endogenous levels of MEK1 phosphorylated at threonine 286. This antibody does not cross-react with phosphorylated MEK2.
Source / Purification
Polyclonal antibodies are produced by immunizing rabbits with a synthetic phospho-peptide (KLH-coupled) corresponding to residues surrounding Thr286 of human MEK1. Antibodies are purified by protein A and peptide affinity chromatography.
Western Blotting
Western blot analysis of extracts from A431, COS and PC12 cells, untreated or nocodazole-treated, using Phospho-MEK1 (Thr286) Antibody (upper) or MEK1 Antibody #9124 (lower).
IP
Immunoprecipitation followed by Western blot analysis of extracts from COS cells, untreated or nocodazole-treated, using Phospho-MEK1 (Thr286) Antibody.
Flow Cytometry
Flow cytometric analysis of untreated Jurkat cells, using Phospho-MEK1 (Thr286) Antibody versus propidium iodide (DNA content). The box indicates Phosho-MEK1 positive cells.
Background
MEK1 and MEK2, also called MAPK or Erk kinases, are dual-specificity protein kinases that function in a mitogen activated protein kinase cascade controlling cell growth and differentiation (1-3). Activation of MEK1 and MEK2 occurs through phosphorylation of two serine residues at positions 217 and 221 (in the activation loop of subdomain VIII) by Raf-like molecules. MEK1/2 is activated by a wide variety of growth factors and cytokines and also by membrane depolarization and calcium influx (1-4). Constitutively active forms of MEK1/2 are sufficient for the transformation of NIH/3T3 cells or the differentiation of PC12 cells (4). MEK activates p44 and p42 MAP kinase by phosphorylating both threonine and tyrosine residues at sites located within the activation loop of kinase subdomain VIII.
MEK1 is phosphorylated at Ser298 by PAK1, which facilitates signal transduction from Raf to MEK1 and Erk2 (5-7). MEK1 is also phosphorylated by cdk5 at Thr286 in mitotic cells, causing negative feedback of the p44/42 MAP kinase pathway (8).
- Crews, C.M. et al. (1992) Science 258, 478-480.
- Alessi, D.R. et al. (1994) EMBO J. 13, 1610-1619.
- Rosen, L.B. et al. (1994) Neuron 12, 1207-1221.
- Cowley, S. et al. (1994) Cell 77, 841-852.
- Xu, B. et al. (1999) J. Biol. Chem. 274, 34029-34035.
- Coles, L.C. and Shaw, P.E. (2002) Oncogene 21, 2236-2244.
- Eblen, S. T. et al. (2002) Mol. Cell. Biol. 22, 6023-6033.
- Sharma, P. et al. (2002) J. Biol. Chem. 277, 528-534.
Application References
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This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.