SignalSilence? PLCγ1 siRNA I (Mouse Specific)

• 产品编号：CST-6413S      品牌：CST       原厂货号：6413S
• 产品分类：DNA和RNA纯化 > RNA干扰 > RNAi Oligos > siRNA
• 应用分类：

描述：

Cell Signaling Technology (CST)公司生产的PLCγ1 siRNA I（小鼠特异的）使得研究者可以使用RNA干扰技术特异性的抑制PLCγ1的表达。这种技术通过将双链RNA分子送入细胞而选择性的沉默某个基因的表达。CST的siRNA产品已经在内部经过严格测试，经western分析确能减少目标蛋白的表达。通过三苯分析对寡核苷酸合成进行逐个碱基的监测，确保了正确的耦联效率。然后用亲和固相萃取纯化寡核苷酸。退火的RNA进一步通过质谱分析，以验证其确切成分。每批都将和以前批次进行质谱比对，以确保批次间最大的一致性。CST公司推荐您在细胞裂解前用100nM PLCγ1 siRNA I（小鼠特异的）进行48-72小时的转染。转染的流程请遵循转染试剂制造商提供的操作流程。使用中如有任何问题，请与CST联系。磷酸肌醇特异性磷脂酶C（PLC）在跨膜信号转导中扮演明显的作用。在对激素、生长因子和神经递质反应的过程中，PLC水解4,5-二磷酸磷酸酰肌醇，产生两个第二信使：1,4,5-三磷酸肌醇（IP3）和甘油二酯（DAG）（1）。至少四个PLC家族已被鉴定，分别为PLC-α、PLC-β、PLC-γ和PLC-δ。磷酸化是调节PLC活性的重要机制之一。PLCγ被受体和非受体酪氨酸激酶激活（2）。PLCγ与EGF和PDGF受体形成复合物，导致PLCγ的Tyr771、783和1245位点磷酸化（3）。Syk对Tyr783的磷酸化激活PLCγ1的酶活性（4）。PLCγ2参与B细胞抗原依赖的信号转导和血小板胶原依赖的信号转导。Btk或Lck对Tyr753、759、1197和1217的磷酸化与PLCγ2的活性相关（5,6）。

1. Singer, W.D. et al. (1997) Annu Rev Biochem 66, 475-509.
2. Margolis, B. et al. (1989) Cell 57, 1101-7.
3. Kim, H.K. et al. (1991) Cell 65, 435-41.
4. Wang, Z. et al. (1998) Mol Cell Biol 18, 590-7.
5. Watanabe, D. et al. (2001) J Biol Chem 276, 38595-601.
6. Ozdener, F. et al. (2002) Mol Pharmacol 62, 672-9.

原厂资料：

Description

SignalSilence® PLCγ1 siRNA I (Mouse Specific) from Cell Signaling Technology (CST) allows the researcher to specifically inhibit PLCγ1 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.

Directions for Use

CST recommends transfection with 100 nM SignalSilence® PLCγ1 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.

Background

Phosphoinositide-specific phospholipase C (PLC) plays a significant role in transmembrane signaling. In response to extracellular stimuli such as hormones, growth factors, and neurotransmitters, PLC hydrolyzes phosphatidylinositol 4,5-bisphosphate (PIP2) to generate two secondary messengers: inositol 1,4,5-triphosphate (IP3) and diacylglycerol (DAG) (1). At least four families of PLCs have been identified: PLCβ, PLCγ, PLCδ, and PLCε. Phosphorylation is one of the key mechanisms that regulate the activity of PLC. PLCγ is activated by both receptor and non-receptor tyrosine kinases (2). PLCγ forms a complex with EGF and PDGF receptors, which leads to the phosphorylation of PLCγ at Tyr771, 783, and 1245 (3). Phosphorylation by Syk at Tyr783 activates the enzymatic activity of PLCγ1 (4). PLCγ2 is engaged in antigen-dependent signaling in B cells and collagen-dependent signaling in platelets. Phosphorylation by Btk or Lck at Tyr753, 759, 1197, and 1217 is correlated with PLCγ2 activity (5,6).

1. Singer, W.D. et al. (1997) Annu Rev Biochem 66, 475-509.
2. Margolis, B. et al. (1989) Cell 57, 1101-7.
3. Kim, H.K. et al. (1991) Cell 65, 435-41.
4. Wang, Z. et al. (1998) Mol Cell Biol 18, 590-7.
5. Watanabe, D. et al. (2001) J Biol Chem 276, 38595-601.
6. Ozdener, F. et al. (2002) Mol Pharmacol 62, 672-9.

注意事项：

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