Product Pathways - MAPK Signaling
PhosphoPlus® MEK1/2 (Ser217/221) Antibody Kit #9120
|9120S||1 Kit (10 western blots)||---||In Stock||---|
|9120||carrier free and custom formulation / quantity||email request|
|Kit Includes||Quantity||Applications||Reactivity||Mol. Wt.||Isotype|
|Phospho-MEK1/2 (Ser217/221) (41G9) Rabbit mAb #9154||100 µl||W, IP||H, M, R, Mk, Dm||45 kDa||Rabbit|
|MEK1/2 (D1A5) Rabbit mAb #8727||100 µl||W, IF-IC, F||H, M, R, Mk, Dm||45 kDa||Rabbit IgG|
|MEK1/2 HeLa Control Cell Extracts #9160||200 µl|
|Anti-rabbit IgG, HRP-linked Antibody #7074||100 µl||W||All||Goat|
|Anti-biotin, HRP-linked Antibody #7075||100 µl||Goat|
|20X LumiGLO® Reagent and 20X Peroxide #7003||5 ml each||W|
|Biotinylated Protein Ladder Detection Pack #7727||100 µl|
Applications Key: W=Western Blotting, IP=Immunoprecipitation, IF-IC=Immunofluorescence (Immunocytochemistry), F=Flow Cytometry
Reactivity Key: H=Human, M=Mouse, R=Rat, Mk=Monkey, Dm=D. melanogaster, All=All Species Expected
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO* is added and emits light during enzyme catalyzed decomposition.
Western blot analysis of extracts from untreated or TPA-treated HeLa and NIH/3T3 cells, using Phospho-MEK1/2 (Ser217/221) (41G9) Rabbit mAb (upper), or MEK1/2 Antibody #9122 (lower).
Western blot analysis of extracts from various cell lines using MEK1/2 (D1A5) Rabbit mAb.
Flow cytometric analysis of Jurkat cells using MEK1/2 (D1A5) Rabbit mAb (blue) compared to Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (red).
Confocal immunofluorescent analysis of HeLa cells using MEK1/2 (D1A5) Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin. Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
The PhosphoPlus® MEK1/2 (Ser217/221) Antibody Kit provides reagents and protocols for the rapid analysis of the phosphorylation status of MEK1/2 at Ser217/221.
Specificity / Sensitivity
Phospho-MEK1/2 (Ser217/221) Antibody detects endogenous levels of MEK1/2 only when phosphorylated at serines 217 and 221. It will also react with MEK1/2 singly phosphorylated at Ser217 and singly phosphorylated at Ser221. MEK1/2 (D1A5) Rabbit mAb recognizes endogenous levels of total MEK1 and MEK2 proteins.
Source / Purification
Monoclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues around Ser217/221 of human MEK1/2 and synthetic peptide corresponding to residues surrounding Ala220 of human MEK1 protein.
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, located 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 PC-12 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.
- Xiao, D. and Singh, S.V. (2002) Cancer Res. 62, 3615-3619. Applications: Western Blotting.
- Watanabe, H. et al. (2001) J. Biol. Chem. 276, 14466-14473. Applications: Western Blotting.
- Jost, M. et al. (2001) J. Biol. Chem. 276, 6320-6326. Applications: Western Blotting.
- DeWindt, L. J. et al. (2000) . Journal of Biological Chemistry 275, 13571-13579. Applications: Western Blotting.
- Soga, S. et al. (1998) . Journal of Biological Chemistry 273, 822-828. Applications: Western Blotting.
- Galbiati, F. et al. (1998) . European Molecular Biology Organization (EMBO) 17, 6633-6648. Applications: Western Blotting.
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This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
U.S. Patent No. 5,675,063.
PhosphoPlus® is a trademark of Cell Signaling Technology, Inc.
LumiGLO® is a registered trademark of Kirkegaard & Perry Laboratories.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.