Cell Signaling Technology

Product Pathways - MAPK Signaling

PhosphoPlus® MEK1/2 (Ser217/221) Antibody Kit #9120

Kit Includes Quantity Applications Reactivity MW (kDa) Isotype
Phospho-MEK1/2 (Ser217/221) (41G9) Rabbit mAb #9154 100 µl W IP H M R Mk Dm (C) 45 Rabbit IgG
MEK1/2 Antibody #9122 100 µl W IP H M R Mk Dm (C) (X) 45 Rabbit
MEK1/2 HeLa Control Cell Extracts #9160 4 mini-blots
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl Goat
Anti-biotin, HRP-linked Antibody #7075 100 µl Goat
20X LumiGLO® Reagent and 20X Peroxide #7003 5 ml
Biotinylated Protein Ladder Detection Pack #7727 100 µl

Applications Key:  W=Western Blotting  IP=Immunoprecipitation
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  C=Chicken  Dm=D. melanogaster  X=Xenopus
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Specificity / Sensitivity

When used in conjunction with our Phototope®-HRP Western Detection System, Phospho-MEK1/2 (Ser217/221) Antibody detects 1 ng phosphorylated MEK1/2 yet will not react with up to 1 µg of nonphosphorylated MEK1/2.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 Antibody detects endogenous levels of total MEK1/2 protein. Neither antibody cross-reacts with related kinases, such as SEK1 (MKK4), MKK3 or MKK6.

Western Blotting

Western Blotting

Western blot analysis of extracts from untreated or TPA-treated HeLa and NIH/3T3 cells, using Phospho-MEK1/2 (Ser217/221) (41G9) Rabbit mAb #9154 (upper), or MEK1/2 Antibody #9122 (lower).

Description

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.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues around Ser217/221 of human MEK1/2. Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues corresponding to the sequence of human MEK1/2 (MEK1/2 Antibody).

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, 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.

  1. Crews, C.M. et al. (1992) Science 258, 478-480.
  2. Alessi, D.R. et al. (1994) EMBO J. 13, 1610-1619.
  3. Rosen, L.B. et al. (1994) Neuron 12, 1207-1221.
  4. Cowley, S. et al. (1994) Cell 77, 841-852.

Application References

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Protocols

Companion Products

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.

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