Cell Signaling Technology

Product Pathways - Jak/Stat Pathway

SignalSilence® PIAS3 siRNA I #9073

No. Size Price
9073S 300 µl ( 3 nmol ) ¥3,224.00 现货查询 购买询价
9073 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,

Homology

Species predicted to react based on 100% sequence homology: Monkey,

Specificity / Sensitivity

SignalSilence® PIAS3 siRNA I inhibits human and monkey PIAS3 expression.

SignalSilence® PIAS3 siRNA I能够抑制人源或猴源PIAS3的表达。

Description

SignalSilence® PIAS3 siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit PIAS3 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.

来自Cell Signaling Technology (CST)的SignalSilence® PIAS3 siRNA I可以帮助研究者通过RNA干扰特异性地抑制PIAS3的表达,这种方法可以通过将双链RNA分子传递到细胞内从而使基因表达有选择的沉默。来自CST的所有的SignalSilence® siRNA产品都是经过内部严格检测的,并且通过Western blot 分析证明确实能够减少目的蛋白的表达。

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.

通过三苯甲基分析每个碱基以监测寡核苷酸的合成,确保合适的配对效率。随后寡核苷酸通过亲和固相萃取法纯化。退火的RNA双链通过质谱分析来证实其精确的组成。每一批产品都通过质谱分析与前面的产品进行比较,来保证不同批次之间的最大一致性。

Western Blotting

Western Blotting

Western blot analysis of extracts from RD cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® PIAS3 siRNA I (+), or SignalSilence® PIAS3 siRNA II #9031 (+), using PIAS3 (D5F9) XP® Rabbit mAb #9042 (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). The PIAS3 (D5F9) XP® Rabbit mAb confirms silencing of PIAS3 expression, while the β-Actin (D6A8) Rabbit mAb is used as a loading control.Western blot分析RD细胞的细胞提取物,转染100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® PIAS3 siRNA I (+), 或 SignalSilence® PIAS3 siRNA II#9031 (+),使用抗体是PIAS3 (D5F9) XP® Rabbit mAb #9042 (上图) 或 β-Actin (D6A8) Rabbit mAb #8457 (下图)。PIAS3 (D5F9) XP® Rabbit mAb证实了PIAS3表达的沉默,而β-Actin (D6A8) Rabbit mAb用来检测内参。

Directions for Use

CST recommends transfection with 100 nM SignalSilence® PIAS3 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.

CST推荐使用100 nM SignalSilence® PIAS3 siRNA I转染,48到72小时后进行细胞裂解。转染步骤按照转染试剂说明书提供的步骤进行。遇到任何使用方面的问题,请随时联系CST。每小瓶可供100次转染,每次转染量相当于在转染24孔板时,每孔总体积为300μl培养基中siRNA的终浓度为100nM。

Background

The protein inhibitor of activated Stat (PIAS) proteins, which include PIAS1, PIAS3, PIASx, and PIASy, were originally characterized based on their interaction with the Stat family of transcription factors (1,2). PIAS1, PIAS3, and PIASx interact with and repress Stat1, Stat3, and Stat4, respectively (1-3). Deletion of PIAS1 leads to inhibition of interferon-inducible genes and increased protection against infection (4). The PIAS family contains a conserved RING domain that has been linked to a function as a small ubiquitin-related modifier (SUMO) ligase, coupling the SUMO conjugating enzyme Ubc9 with its substrate proteins (5,6). Numerous studies have now shown that PIAS family members can regulate the activity of transcription factors through distinct mechanisms, including NF-κB (7,8), c-Jun, p53 (5,9), Oct-4 (10), and Smads (11,12). The activity of PIAS1 is regulated by both phosphorylation and arginine methylation. Inflammatory stimuli can induce IKK-mediated phosphorylation of PIAS1 at Ser90, which is required for its activity (13). In addition, PRMT1 induces arginine methylation of PIAS1 at Arg303 following interferon treatment and is associated with its repressive activity on Stat1 (14).

PIAS (激活的Stat的蛋白抑制剂)蛋白, 包括PIAS1, PIAS3, PIASx 和 PIASy,他们最初是通过与Stat转录因子的相互作用而被鉴定(1,2)。PIAS1, PIAS3 和 PIASx分别与Stat1, Stat3 和 Stat4相互作用并抑制他们(1-3)。PIAS1的缺失导致了对干扰素诱导的基因的抑制并增强对感染的抵抗(4)。PIAS家族的成员包含一个保守的RING区域,此区域的功能类似于 SUMO (小泛素相关修饰)连接酶, 偶联SUMO结合酶Ubc9及其底物蛋白(5,6)。大量的研究表明PIAS家族的成员能够通过不同的机制调控转录因子的活性,包括NF-κB (7,8), c-Jun, p53 (5,9), Oct-4 (11)和 Smads (11,12)。PIAS1的活性是通过磷酸化和精氨酸的甲基化调节的。炎症刺激能够诱导IKK介导PIAS1在Ser90位点的磷酸化,这对其激活是必需的(13)。另外,在干扰素的处理下PRMT1诱导PIAS1在Arg303处的甲基化,这与其抑制Stat1活性相关(14)。

  1. Liu, B. et al. (1998) Proc Natl Acad Sci USA 95, 10626-31.
  2. Chung, C.D. et al. (1997) Science 278, 1803-5.
  3. Arora, T. et al. (2003) J Biol Chem 278, 21327-30.
  4. Liu, B. et al. (2004) Nat Immunol 5, 891-8.
  5. Schmidt, D. and Müller, S. (2002) Proc Natl Acad Sci USA 99, 2872-7.
  6. Kotaja, N. et al. (2002) Mol Cell Biol 22, 5222-34.
  7. Liu, B. et al. (2005) Mol Cell Biol 25, 1113-23.
  8. Tahk, S. et al. (2007) Proc Natl Acad Sci USA 104, 11643-8.
  9. Bischof, O. et al. (2006) Mol Cell 22, 783-94.
  10. Tolkunova, E. et al. (2007) J Mol Biol 374, 1200-12.
  11. Long, J. et al. (2004) Proc Natl Acad Sci USA 101, 99-104.
  12. Murdoch, R.N. and Edwards, T. (1992) Biochem Int 28, 1029-37.
  13. Liu, B. et al. (2007) Cell 129, 903-14.
  14. Weber, S. et al. (2009) Genes Dev 23, 118-32.

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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.

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

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