|6243||SignalSilence® p38 MAPK siRNA II||
Fluorescent detection of SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 in living HeLa cells 24 hours post-transfection, demonstrating nearly 100% transfection efficiency.
Western blot analysis of extracts from HeLa cells transfected with 20 nM control siRNA #6201, or 5, 20 nM pool p38 MAP Kianse siRNA, using p38 MAP Kinase Antibody #9212 (upper), p44/42 MAP Kinase Antibody #9102 (middle) and SAPK/JNK (56G8) Rabbit mAb #9258 (lower) in combination with ATP-Citrate Lyase Antibody #4332. The p38 MAP Kinase Antibody confirms silencing of p38 MAP Kinase expression, and ATP-Citrate Lyase Antibody is used to control for loading and specificity of pool p38 MAP Kinase siRNA. The experiment also demonstrates specifically that the pool p38 MAPK siRNA does not interfere with the expression of p44/42 MAPK and SAPK/JNK.
CST recommends transfection with 5 to 20 nM pool p38 MAP Kinase siRNA 48 hours prior to cell lysis. For transfection procedure, follow protocol provided by the transfection reagent manufacturer. Please feel free to contact CST with any questions on use.
SignalSilence® siRNA is supplied in RNAse-free water. Aliquot and store at -20ºC.
SignalSilence® Pool p38 MAP Kinase siRNA from Cell Signaling Technology allows the researcher to specifically inhibit p38 MAP Kinase expression using RNA interference, a method in which gene expression can be selectively silenced through the delivery of double stranded RNA molecules into the cell. The p38 MAP Kinase siRNA is a hetergeneous mixture of 21-22 bp siRNAs. A 418 bp cDNA template of mouse p38 MAP Kinase was transcribed by T7 RNA polymerase to create double-stranded RNA (dsRNA). RNase III was used to cleave the dsRNA in the presence of Mn2+ buffer to 21-22 bp siRNA. All SignalSilence® siRNA products are rigorously tested in-house and have been shown to reduce protein expression in specified cell lines.
p38 MAP kinase (MAPK), also called RK (1) or CSBP (2), is the mammalian orthologue of the yeast HOG kinase that participates in a signaling cascade controlling cellular responses to cytokines and stress (1-4). Four isoforms of p38 MAPK, p38α, β, γ (also known as Erk6 or SAPK3), and δ (also known as SAPK4) have been identified. Similar to the SAPK/JNK pathway, p38 MAPK is activated by a variety of cellular stresses including osmotic shock, inflammatory cytokines, lipopolysaccharide (LPS), UV light, and growth factors (1-5). MKK3, MKK6, and SEK activate p38 MAPK by phosphorylation at Thr180 and Tyr182. Activated p38 MAPK has been shown to phosphorylate and activate MAPKAP kinase 2 (3) and to phosphorylate the transcription factors ATF-2 (5), Max (6), and MEF2 (5-8). SB203580 (4-(4-fluorophenyl)-2-(4-methylsulfinylphenyl)-5-(4-pyridyl)-imidazole) is a selective inhibitor of p38 MAPK. This compound inhibits the activation of MAPKAPK-2 by p38 MAPK and subsequent phosphorylation of HSP27 (9). SB203580 inhibits p38 MAPK catalytic activity by binding to the ATP-binding pocket, but does not inhibit phosphorylation of p38 MAPK by upstream kinases (10).
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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.