Product Pathways - MAPK Signaling
SignalSilence® Pool p42 MAPK siRNA #6391
Western Blotting
Western blot analysis of extracts from HeLa cells transfected with 20 nM control siRNA #6201 (-) or 2, 5, or 20 nM pool 42 MAPK siRNA, using p44/42 MAP Kinase Antibody #9102 (upper), p38 MAP Kinase Antibody #9212 (middle) and SAPK/JNK (56G8) Rabbit mAb #9258 (lower) in combination with ATP-Citrate Lyase Antibody #4332. The 44/42 MAP Kinase Antibody confirms silencing of p42 MAP Kinase expression, and ATP-Citrate Lyase Antibody is used to control for loading and specificity of pool p42 MAP Kinase siRNA. The experiment also demonstrates specifically that the pool p42 MAPK siRNA does not interfere with the expression of p44 MAPK, p38 MAPK or SAPK/JNKs.
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
Background
Mitogen-activated protein kinases (MAPKs) are a widely conserved family of serine/threonine protein kinases involved in many cellular programs such as cell proliferation, differentiation, motility, and death. The p44/42 MAPK (ERK1/2) signaling pathway can be activated in response to a diverse range of extracellular stimuli including mitogens, growth factors, and cytokines (1-3) and is an important target in the diagnosis and treatment of cancer (4). Upon stimulation, a sequential three-part protein kinase cascade is initiated, consisting of a MAP kinase kinase kinase (MAPKKK or MAP3K), a MAP kinase kinase (MAPKK or MAP2K), and a MAP kinase (MAPK). Multiple p44/42 MAP3Ks have been identified, including members of the Raf family as well as Mos and Tpl2/Cot. MEK1 and MEK2 are the primary MAPKKs in this pathway (5,6). MEK1 and MEK2 activate p44 and p42 through phosphorylation of activation loop residues Thr202/Tyr204 and Thr185/Tyr187, respectively. Several downstream targets of p44/42 have been identified, including p90RSK (7) and the transcription factor Elk-1 (8,9). p44/42 are negatively regulated by a family of dual-specificity (Thr/Tyr) MAPK phosphatases, known as DUSPs or MKPs (10), along with MEK inhibitors such as U0126 and PD98059.
- Roux, P.P. and Blenis, J. (2004) Microbiol Mol Biol Rev 68, 320-44.
- Baccarini, M. (2005) FEBS Lett 579, 3271-7.
- Meloche, S. and Pouyssegur, J. (2007) Oncogene 26, 3227-39.
- Roberts, P.J. and Der, C.J. (2007) Oncogene 26, 3291-310.
- Rubinfeld, H. and Seger, R. (2005) Mol Biotechnol 31, 151-74.
- Murphy, L.O. and Blenis, J. (2006) Trends Biochem Sci 31, 268-75.
- Dalby, K.N. et al. (1998) J Biol Chem 273, 1496-505.
- Marais, R. et al. (1993) Cell 73, 381-93.
- Kortenjann, M. et al. (1994) Mol Cell Biol 14, 4815-24.
- Owens, D.M. and Keyse, S.M. (2007) Oncogene 26, 3203-13.
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
- 6390 SignalSilence® Pool p42 MAPK siRNA Kit
- 9102 p44/42 MAP Kinase Antibody
- 9101 Phospho-p44/42 MAPK (Thr202/Tyr204) Antibody
- 3372 Erk5 Antibody
- 6435 SignalSilence® p44 MAPK siRNA Kit (Human Specific)
- 6436 SignalSilence® p44 MAPK siRNA (Human Specific)
- 6425 SignalSilence® MEK1 siRNA Kit (Human Specific)
- 6426 SignalSilence® MEK1 siRNA (Human Specific)
- 6430 SignalSilence® MEK2 siRNA Kit (Human Specific)
- 6431 SignalSilence® MEK2 siRNA (Human Specific)
- 6201 SignalSilence® Control siRNA (Fluorescein Conjugate)
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.
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.
