Upstream / Downstream

pathwayImage

Explore pathways related to this product.

Important Ordering Details

Custom Ordering Details: Product is assembled upon order. Please allow up to three business days for your product to be processed.

Antibody Guarantee

CST Antibody Performance Guarantee

LEARN MORE  

To Purchase # 12175S

12175S 300 µl (3 nmol)

To get local purchase information on this product, click here

Questions?

Find answers on our FAQs page.

ANSWERS  

Visit PhosphoSitePlus®

PTM information and tools available.

LEARN MORE

REACTIVITY
H

Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® EWS siRNA I (+), or SignalSilence® EWS siRNA II #12216 (+), using EWS Antibody #11910 (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). The EWS Antibody confirms silencing of EWS expression, while the β-Actin (D6A8) Rabbit mAb is used as a loading control.

Learn more about how we get our images
Image

Product Usage Information

CST recommends transfection with 100 nM SignalSilence® EWS 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.


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

Product Description

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

The Ewing sarcoma (EWS) protein is a member of the multifunctional FET (FUS, EWS, and TAF15) family of proteins (1,2). These proteins are RNA and DNA binding proteins that are thought to be important for both transcriptional regulation and RNA processing. EWS can be found as part of a fusion protein with various E-twenty six (ETS) family transcription factors, most commonly Fli-1, in the Ewing sarcoma family of tumors (1-4). The amino terminus of the EWS protein, containing the transcriptional activation domain, is fused to the DNA binding domain of the ETS transcription factor, causing aberrant expression of target genes (1-5). EWS interacts with the transcription initiation complex via TFIID and RNA polymerase II subunits, as well as transcriptional regulators, such as Brn3A and CBP/p300, which suggests a role for EWS in transcriptional regulation (1,6-9). EWS also interacts with multiple components of the splicing machinery, implicating a role for EWS in RNA processing (1,10-12). EWS regulates the expression of cyclin D1, which controls G1-S phase transition during the cell cycle, at the level of transcriptional activation and mRNA splicing. The EWS-Fli-1 fusion protein has been shown to promote the expression of the cyclin D1b splice variant in Ewing sarcoma cells (13). In addition, EWS regulates the DNA damage-induced alternative splicing of genes involved in DNA repair and stress response and is required for cell viability upon DNA damage (14). Consistent with these results, EWS knockout mice display hypersensitivity to ionizing radiation and premature cellular senescence, suggesting a role for EWS in homologous recombination and maintenance of genomic stability (15).


1.  Law, W.J. et al. (2006) Brief Funct Genomic Proteomic 5, 8-14.

2.  Kovar, H. (2011) Sarcoma 2011, 837474.

3.  Delattre, O. et al. (1992) Nature 359, 162-5.

4.  May, W.A. et al. (1993) Mol Cell Biol 13, 7393-8.

5.  Bertolotti, A. et al. (1996) EMBO J 15, 5022-31.

6.  Sorensen, P.H. et al. (1994) Nat Genet 6, 146-51.

7.  Bertolotti, A. et al. (1998) Mol Cell Biol 18, 1489-97.

8.  Araya, N. et al. (2003) J Biol Chem 278, 5427-32.

9.  Thomas, G.R. and Latchman, D.S. Cancer Biol Ther 1, 428-32.

10.  Chansky, H.A. et al. (2001) Cancer Res 61, 3586-90.

11.  Yang, L. et al. (2000) J Biol Chem 275, 37612-8.

12.  Knoop, L.L. and Baker, S.J. (2001) J Biol Chem 276, 22317-22.

13.  Sanchez, G. et al. (2008) Proc Natl Acad Sci U S A 105, 6004-9.

14.  Paronetto, M.P. et al. (2011) Mol Cell 43, 353-68.

15.  Li, H. et al. (2007) J Clin Invest 117, 1314-23.


Entrez-Gene Id 2130
Swiss-Prot Acc. Q01844


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