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13553
SignalSilence® Sara siRNA II
siRNA
siRNA

SignalSilence® Sara siRNA II #13553

Citations (0)
SignalSilence® Sara siRNA II: Image 1

Western blot analysis of extracts from SH-SY-5Y cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® Sara siRNA I #13542 (+) or SignalSilence® Sara siRNA II (+), 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.

Supporting Data

REACTIVITY

Application Key:

  • W-Western
  • IP-Immunoprecipitation
  • IHC-Immunohistochemistry
  • ChIP-Chromatin Immunoprecipitation
  • IF-Immunofluorescence
  • F-Flow Cytometry
  • E-P-ELISA-Peptide

Species Cross-Reactivity Key:

  • H-Human
  • M-Mouse
  • R-Rat
  • Hm-Hamster
  • Mk-Monkey
  • Mi-Mink
  • C-Chicken
  • Dm-D. melanogaster
  • X-Xenopus
  • Z-Zebrafish
  • B-Bovine
  • Dg-Dog
  • Pg-Pig
  • Sc-S. cerevisiae
  • Ce-C. elegans
  • Hr-Horse
  • All-All Species Expected

Product Usage Information

CST recommends transfection with 100 nM SignalSilence® Sara siRNA II 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.

Storage

SignalSilence® siRNA is supplied in RNAse-free water. Aliquot and store at -20ºC.

Product Description

SignalSilence® Sara siRNA II 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.

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.

Pathways & Proteins

Explore pathways + proteins related to this product.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST's products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST's Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

For Research Use Only. Not For Use In Diagnostic Procedures.
CST is a trademark of Cell Signaling Technology, Inc.
SignalSilence is a registered trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
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