Cell Signaling Technology

Product Pathways - TGF-beta/Smad Signaling

SignalSilence® Sara siRNA I #13542

No. Size Price
13542S 300 µl ( 3 nmol ) ¥3,228.00 现货查询 购买询价
13542 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
TFN Human,

Species cross-reactivity is determined by western blot.

Applications Key: TFN=Transfection,

Description

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

Western Blotting

Western Blotting

Western blot analysis of extracts from SH-SY5Y cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® Sara siRNA I (+) or SignalSilence® Sara siRNA II #13553 (+), using Sara (D5X4F) Rabbit mAb #13285 (upper) or α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower). The Sara (D5X4F) Rabbit mAb confirms silencing of Sara expression, while the α-Actinin (D6F6) XP® Rabbit mAb is used as a loading control.

Directions for Use

CST recommends transfection with 100 nM SignalSilence® Sara siRNA I 48 to 72 hours prior to cell lysis. For transfection procedure, follow the 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

The Smad anchor for receptor activation (SARA, ZFYVE9) protein is an FYVE domain-containing protein originally identified as a regulator of TGF-β signaling (1). FYVE domains are zinc finger-like domains that bind to phosphatidylinositol 3-phosphate and are responsible for endosomal trafficking (2). While the role of Sara in TGF-β signaling has been questioned (3,4), early research studies demonstrate that Sara enhances TGF-β signaling by binding and recruiting non-activated Smad2 and Smad3 to the TGF-β receptor complex (1). Upon Smad2 activation, Sara dissociates from the complex while phosphorylated Smad2/3 translocates to the nucleus to bind to the common Smad, Smad4. Sara can also function as an anchor for the protein phosphatase 1 (PP1c) catalytic subunit, which is involved in the Smad7-mediated dephosphorylation of TGF-β type I receptor (5,6). Additional research studies show that expression of Sara plays a critical role in maintenance of the epithelial cell phenotype and that expression is regulated during the epithelial-to-mesenchymal transition (EMT) and fibrosis (7,8).

  1. Tsukazaki, T. et al. (1998) Cell 95, 779-91.
  2. Itoh, F. et al. (2002) Genes Cells 7, 321-31.
  3. Bakkebø, M. et al. (2012) FEBS Lett 586, 3367-72.
  4. Goto, D. et al. (2001) Biochem Biophys Res Commun 281, 1100-5.
  5. Bennett, D. and Alphey, L. (2002) Nat Genet 31, 419-23.
  6. Shi, W. et al. (2004) J Cell Biol 164, 291-300.
  7. Runyan, C.E. et al. (2009) J Biol Chem 284, 25181-9.
  8. Zhao, B.M. and Hoffmann, F.M. (2006) Mol Biol Cell 17, 3819-31.

Application References

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Protocols

Companion Products


For Research Use Only. Not For Use In Diagnostic Procedures.

XP is a registered trademark of Cell Signaling Technology, Inc.

SignalSilence is a registered trademark of Cell Signaling Technology, Inc.

CST is a trademark of Cell Signaling Technology, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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