Product Pathways - Chromatin Regulation / Epigenetics
SignalSilence® SP1 siRNA I #12104
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 HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® SP1 siRNA I (+), or SignalSilence® SP1 siRNA II #12106 (+), using SP1 Antibody #5931 (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). The SP1 Antibody confirms silencing of SP1 expression, while the β-Actin (D6A8) Rabbit mAb is used as a loading control. The SP1 Antibody cross-reacts with a protein of unknown origin at 55 kDa, which is unaffected by SignalSilence® SP1 siRNA I or SignalSilence® SP1 siRNA II #12106.
SignalSilence® SP1 siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit SP1 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 SignalSilence® SP1 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.
Specificity protein 1 (SP1) is a ubiquitously expressed transcription factor belonging to the family of C2H2-type zinc finger containing DNA-binding proteins. SP1 binds GC-rich motifs with high affinity and regulates the expression of numerous mammalian genes (1,2). It interacts with many other transcription factors, such as c-Myc, EGR1, and Stat1, and with basal transcription machinery components. SP1 interacts with chromatin-modifying factors, such as histone deacetylases (HDACs) and p300 in chromatin remodeling. Transcriptional activity and stability of SP1 are regulated by post-translational modification, including phosphorylation, acetylation, ubiquitination, and glycosylation (3). Glycosylation of SP1 following insulin treatment leads to increased nuclear localization, while glucagon treatment increases cytoplasmic SP1 levels (4-6). Investigators have found high levels of SP1 in patients with Alzheimer's disease (7).
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- Song, J. et al. (2003) Int J Mol Med 11, 547-53.
- Tan, N.Y. and Khachigian, L.M. (2009) Mol Cell Biol 29, 2483-8.
- Majumdar, G. et al. (2003) Am J Physiol Endocrinol Metab 285, E584-91.
- Majumdar, G. et al. (2006) J Biol Chem 281, 3642-50.
- Solomon, S.S. et al. (2008) Life Sci 83, 305-12.
- Citron, B.A. et al. (2008) J Neurosci Res 86, 2499-504.
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- 6568 SignalSilence® Control siRNA (Unconjugated)
- 6201 SignalSilence® Control siRNA (Fluorescein Conjugate)
- 12106 SignalSilence® SP1 siRNA II
- 5931 SP1 Antibody
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.