Cell Signaling Technology

Product Pathways - MAPK Signaling

SignalSilence® MEK2 siRNA (Human Specific) #6431

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells transfected with 100 nM control siRNA #6201 (lane 1), MEK1 siRNA (lane 2), MEK2 siRNA (lane 3) or MEK1 and MEK2 siRNA (Lane 4), using MEK2 Antibody #9125 (upper) or MEK1 Antibody #9124 (lower) in combination with GCK Antibody #3782. The MEK2 Antibody confirms silencing of MEK2 expression, MEK1 Antibody demonstrates MEK2 siRNA has no effect on its homologue MEK1, and GCK Antibody is used to control for loading and specificity of MEK2 siRNA.

Fluorescent Detection

Fluorescent Detection

Fluorescent detection of SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 in living HeLa cells 24 hours post-transfection, demonstrating nearly 100% transfection efficiency.

Directions for Use

CST recommends transfection with 100nM human-specific MEK2 siRNA 48 to 72 hours prior to cell lysis. See Protocol for transfection procedure.

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.

Small interfering RNA (siRNA) has been used to specifically silence MEK2 expression in HEK293T cells (5).

  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.
  5. Castro-Obregon, S. et al. (2004) J. Biol. Chem. In press, .

Application References

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Companion Products

Limited Use Label License, RNA interference: This product is licensed under European Patent 1144623 and foreign equivalents from Ribopharma AG, Kulmbach, Germany and is provided only for use in non-commercial research specifically excluding use (a) in drug discovery or drug development, including target identification or target validation, by or on behalf of a commercial entity, (b) for contract research or commercial screening services, (c) for the production or manufacture of siRNA-related products for sale, or (d) for the generation of commercial databases for sale to Third Parties. Information about licenses for these and other commercial uses is available from Ribopharma AG, Fritz-Hornschuch-Str. 9, D-95326 Kulmbach, Germany.

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