Product Pathways - MAPK Signaling
SignalSilence® MEK1 siRNA (Human Specific) #6426
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 MEK1 Antibody #9124 (upper) or MEK2 Antibody #9125 (lower) in combination with GCK Antibody #3782. The MEK1 Antibody confirms silencing of MEK1 expression, MEK2 Antibody demonstrates MEK1 siRNA has no effect on its homologue MEK2, and GCK Antibody is used to control for loading and specificity of MEK1 siRNA.
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 MEK1 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 MEK1 expression in HEK293T cells (5).
- 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.
- Castro-Obregon, S. et al. (2004) J. Biol. Chem. In press, .
Application References
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
Companion Products
- 6430 SignalSilence® MEK2 siRNA Kit (Human Specific)
- 6431 SignalSilence® MEK2 siRNA (Human Specific)
- 9122 MEK1/2 Antibody
- 9124 MEK1 Antibody
- 9125 MEK2 Antibody
- 9146 MEK1 (30C8) Rabbit mAb
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.
