Cell Signaling Technology

Product Pathways - Ca / cAMP / Lipid Signaling

SignalSilence® Gelsolin siRNA I #6469

Applications Reactivity
Transfection H

Reactivity Key:  H=Human
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® Gelsolin siRNA I (+), or SignalSilence® Gelsolin siRNA II #6473 (+) using Gelsolin Antibody #8090 (upper) or α-Tubulin (11H10) Rabbit mAb #2125 (lower). The Gelsolin Antibody confirms silencing of gelsolin expression, while the α-Tubulin (11H10) Rabbit mAb is used as a loading control.

Description

SignalSilence® Gelsolin siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit gelsolin 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.

Quality Control

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® Gelsolin 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.

Background

Gelsolin (actin-depolymerizing factor, ADF, AGEL, Brevin) is an 83 kDa protein that shares structural and functional homology to villin and adseverin/scinderin (1,2). Gelsolin plays an important role in actin filament assembly by capping and severing actin proteins in a Ca2+-dependent manner (3,4). Gelsolin is important for cellular events (e.g., membrane ruffling, chemotaxis, ciliogenesis) that require cytoskeletal remodeling (3); accordingly, cells from gelsolin knockout mice exhibit motility defects, including a failure to ruffle in response to growth factor stimulation (5,6). In humans, defects in gelsolin have been linked to amyloidosis type 5 (AMYL5), a hereditary disease characterized by cranial neuropathy, which appears to result from gelsolin amyloid deposition (7).

  1. Vitale, M.L. et al. (1991) J Cell Biol 113, 1057-67.
  2. Lueck, A. et al. (1998) J Cell Sci 111 ( Pt 24), 3633-43.
  3. Sun, H.Q. et al. (1999) J Biol Chem 274, 33179-82.
  4. Li, G.H. et al. (2010) Med Res Rev, Epub ahead of print.
  5. Azuma, T. et al. (1998) EMBO J 17, 1362-70.
  6. Lu, M. et al. (1997) J Cell Biol 138, 1279-87.
  7. Haltia, M. et al. (1990) Biochem Biophys Res Commun 167, 927-32.

Application References

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Companion Products

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.

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