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6346S 300 µl (3 nmol) $249.00


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Western blot analysis of extracts from NIH/3T3 cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® IRS-1 siRNA I (Mouse Specific) (+), or SignalSilence® IRS-1 siRNA II (Mouse Specific) #6374 (+), using IRS-1 (59G8) Rabbit mAb #2390 (upper) or α-Tubulin (11H10) Rabbit mAb #2125 (lower). The IRS-1 (59G8) Rabbit mAb confirms silencing of IRS-1 expression, while the α-Tubulin (11H10) Rabbit mAb is used as a loading control.

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Product Usage Information

CST recommends transfection with 100 nM SignalSilence® IRS-1 siRNA I (Mouse Specific) 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® IRS-1 siRNA I (Mouse Specific) from Cell Signaling Technology (CST) allows the researcher to specifically inhibit IRS-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.

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.

Insulin receptor substrate 1 (IRS-1) is one of the major substrates of the insulin receptor kinase (1). IRS-1 contains multiple tyrosine phosphorylation motifs that serve as docking sites for SH2-domain containing proteins that mediate the metabolic and growth-promoting functions of insulin (2-4). IRS-1 also contains over 30 potential serine/threonine phosphorylation sites. Ser307 of IRS-1 is phosphorylated by JNK (5) and IKK (6) while Ser789 is phosphorylated by SIK-2, a member of the AMPK family (7). The PKC and mTOR pathways mediate phosphorylation of IRS-1 at Ser612 and Ser636/639, respectively (8,9). Phosphorylation of IRS-1 at Ser1101 is mediated by PKCθ and results in an inhibition of insulin signaling in the cell, suggesting a potential mechanism for insulin resistance in some models of obesity (10).

1.  Sun, X.J. et al. (1991) Nature 352, 73-77.

2.  Sun, X.J. et al. (1992) J. Biol. Chem. 267, 22662-22672.

3.  Myers Jr., M.G. et al. (1993) Endocrinology 132, 1421-1430.

4.  Wang, L.M. et al. (1993) Science 261, 1591-1594.

5.  Rui, L. et al. (1997) J. Clin. Invest. 107, 181-189.

6.  Gao, Z. et al. (2002) J. Biol. Chem. 277, 48115-48121.

7.  Horike, N. et al. (2003) J. Biol. Chem. 278, 18440-18447.

8.  Ozes, O.N. et al. (2001) Proc. Natl. Acad. Sci. USA 98, 4640-4645.

9.  De Fea, K. and Ruth, R.A. (1997) Biochemistry 36, 12939-12947.

10.  Li, Y. et al. (2004) J. Biol. Chem. 279, 45304-45307.

Entrez-Gene Id 16367
Swiss-Prot Acc. P35569

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

SignalSilence® IRS-1 siRNA I (Mouse Specific)