Product Pathways - Tyrosine Kinase / Adaptors
SignalSilence® β-Arrestin 1 siRNA I #6218
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 (-) or SignalSilence® β-Arrestin 1 siRNA I (+) using β-Arrestin 2 (C16D9) Rabbit mAb #3857 (upper) or α-Tubulin (11H10) Rabbit mAb #2125 (lower). The β-Arrestin 2 (C16D9) Rabbit mAb confirms specificity of β-Arrestin 1 siRNA I, while the α-Tubulin (11H10) Rabbit mAb is used as a loading control.
Western blot analysis of extracts from HeLa cells transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-) or SignalSilence® β-Arrestin 1 siRNA I (+) using β-Arrestin 1/2 (D24H9) XP® Rabbit mAb #4674 (upper) or α-Tubulin (11H10) Rabbit mAb #2125 (lower). The β-Arrestin 1/2 (D24H9) Rabbit mAb confirms silencing of β-Arrestin expression, while the α-Tubulin (11H10) Rabbit mAb is used to control for loading and specificity of β-Arrestin 1 siRNA.
SignalSilence® β-Arrestin 1 siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit β-arrestin 1 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 β-Arrestin 1 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.
Arrestin proteins function as negative regulators of G protein-coupled receptor (GPCR) signaling. Cognate ligand binding stimulates GPCR phosphorylation, which is followed by binding of arrestin to the phosphorylated GPCR and the eventual internalization of the receptor and desensitization of GPCR signaling (1). Four distinct mammalian arrestin proteins are known. Arrestin 1 (also known as S-arrestin) and arrestin 4 (X-arrestin) are localized to retinal rods and cones, respectively. Arrestin 2 (also known as β-arrestin 1) and arrestin 3 (β-arrestin 2) are ubiquitously expressed and bind to most GPCRs (2). β-arrestins function as adaptor and scaffold proteins and play important roles in other processes, such as recruiting c-Src family proteins to GPCRs in Erk activation pathways (3,4). β-arrestins are also involved in some receptor tyrosine kinase signaling pathways (5-8). Additional evidence suggests that β-arrestins translocate to the nucleus and help regulate transcription by binding transcriptional cofactors (9,10).
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- Lefkowitz, R.J. and Whalen, E.J. (2004) Curr Opin Cell Biol 16, 162-8.
- Waters, C.M. et al. (2005) Cell Signal 17, 263-77.
- Kang, J. et al. (2005) Cell 123, 833-47.
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- 6568 SignalSilence® Control siRNA (Unconjugated)
- 6201 SignalSilence® Control siRNA (Fluorescein Conjugate)
- 4674 β-Arrestin 1/2 (D24H9) 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.