Product Pathways - MAPK Signaling
SignalSilence® ATF-2 siRNA I #6433
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Western blot analysis of extracts from ACHN cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® ATF-2 siRNA I (+) or SignalSilence® ATF-2 siRNA II #6443 (+), using ATF-2 (20F1) Rabbit mAb #9226 (upper) or α-Tubulin (11H10) Rabbit mAb #2125 (lower). The ATF-2 (20F1) Rabbit mAb confirms silencing of ATF-2 expression, while the α-Tubulin (11H10) Rabbit mAb is used as a loading control.
SignalSilence® ATF-2 siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit ATF-2 expression using RNA interference, a method whereby gene expression can be selectively silenced through the delivery of double stranded RNA molecules into the cell. All SignalSilence® siRNA products from CST are rigorously tested in-house and have been shown to reduce target protein expression by western analysis.
Oligonucleotide synthesis is monitored base by base through trityl analysis to ensure appropriate coupling efficiency. The oligo is subsequently purified by affinity-solid phase extraction. The annealed RNA duplex is further analyzed by mass spectrometry to verify the exact composition of the duplex. Each lot is compared to the previous lot by mass spectrometry to ensure maximum lot-to-lot consistency.
Directions for Use
CST recommends transfection with 100 nM ATF-2 siRNA I 48 to 72 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.
Each vial contains the equivalent of 100 transfections, which corresponds to a final siRNA concentration of 100 nM per transfection in a 24-well plate with a total volume of 300 μl per well.
The transcription factor ATF-2 (also called CRE-BP1) binds to both AP-1 and CRE DNA response elements and is a member of the ATF/CREB family of leucine zipper proteins (1). ATF-2 interacts with a variety of viral oncoproteins and cellular tumor suppressors and is a target of the SAPK/JNK and p38 MAP kinase signaling pathways (2-4). Various forms of cellular stress, including genotoxic agents, inflammatory cytokines, and UV irradiation, stimulate the transcriptional activity of ATF-2. Cellular stress activates ATF-2 by phosphorylation of Thr69 and Thr71 (2-4). Both SAPK and p38 MAPK have been shown to phosphorylate ATF-2 at these sites in vitro and in cells transfected with ATF-2. Mutations of these sites result in the loss of stress-induced transcription by ATF-2 (2-4). In addition, mutations at these sites reduce the ability of E1A and Rb to stimulate gene expression via ATF-2 (2).
- Abdel-Hafiz, H.A. et al. (1992) Mol. Endocrinol. 6, 2079-2089.
- Gupta, S. et al. (1995) Science 267, 389-393.
- van Dam, H. et al. (1995) EMBO J. 14, 1798-1811.
- Livingstone, C. et al. (1995) EMBO J. 14, 1785-1797.
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- 6568 SignalSilence® Control siRNA (Unconjugated)
- 6201 SignalSilence® Control siRNA (Fluorescein Conjugate)
- 6443 SignalSilence® ATF-2 siRNA II
- 9226 ATF-2 (20F1) 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.
For Research Use Only. Not For Use In Diagnostic Procedures.