SignalSilence® ADAM9 siRNA I可以抑制人和猴ADAM9的表达。CST的基因沉默ADAM9 siRNA I 允许研究者采用RNA干扰的方法来特异性地抑制ADAM9的表达，这是一种通过去除细胞内双链RNA分子而使基因表达能够选择性沉默的方法。所有CST的基因沉默siRNA产品都经过内部的严格检测并且采用western方法证实可以减少蛋白表达。CST推荐细胞裂解前采用100 nM SignalSilence® ADAM9 siRNA I 作用48-72小时。对于每一个转染步骤，遵循转染试剂商提供的产品使用步骤。如果使用中有任何问题，可与CST联系。每一个产品包含了100次转染需要的量，相当于每次转染siRNA的终浓度为100nM，24孔板的每孔总体积为300微升。ADAM（解聚素和金属蛋白酶）家族具有多种结构域的膜蛋白，通过使细胞表面蛋白脱落例如细胞因子和生长因子，通过影响细胞粘附分子与细胞外基质的作用，以及通过直接重塑细胞外基质，从而影响细胞信号转导和粘附。ADAM家族成员保守的结构域包括一个功能前区，一个锌依赖的金属蛋白酶结构域，一个解聚素结构域，一个富含半胱氨酸的结构域，一个表皮生长因子样序列以及一个胞质尾区(1,2)。该功能前区可以辅助蛋白折叠。解聚素和富含半胱氨酸的结构域通过与整合素结合，至少部分介导折叠。胞质尾区的磷酸化以及它与其它信号转导蛋白的相互作用可能影响细胞内和细胞外的信号转导(1)。ADAM9广泛分布，并已被证实可以影响皮肤角蛋白的迁移(3,4)。研究表明ADAM9在前列腺癌(5)、胰腺癌(6)、胃癌(7)中过度表达，并且与小细胞肺癌的浸润和转移(8)有关。研究表明ADAM9的剪接片段(50 kDa)形式具有蛋白酶活性与肿瘤细胞浸润有关(9)。
原厂资料：
Homology
Species predicted to react based on 100% sequence homology:Monkey
Specificity / Sensitivity
SignalSilence® ADAM9 siRNA I inhibits human and monkey ADAM9 expression.
Description
SignalSilence® ADAM9 siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit ADAM9 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® ADAM9 siRNA I 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.
ackground
The ADAM (A Disintegrin and A Metalloprotease) family of multidomain membrane proteins influences cell signaling and adhesion by shedding cell surface proteins such as cytokines and growth factors, by influencing cell adhesion to the extracellular matrix (ECM), and by directly remodeling the ECM. Conserved domains in ADAM family members include a prodomain, a zinc-dependent metalloprotease domain, a disintegrin domain, a cysteine-rich domain, an EGF-like sequence, and a short cytoplasmic tail (1,2). The prodomain is thought to aid in protein folding. Disintegrin and cysteine-rich domains mediate adhesion, at least in part, through binding to integrins. Phosphorylation of the cytoplasmic tail as well as its interaction with other signaling proteins may influence intra- and extracellular signaling (1). ADAM9 is widely distributed and has been shown to affect migration in skin keratinocytes (3,4). Research studies have shown that ADAM9 is overexpressed in prostate cancer (5), pancreatic cancer (6), gastric cancer (7), and has been linked to invasion and metastasis in small cell lung cancer (8). Research has also shown that an alternatively spliced short (50 kDa) form of ADAM9 containing protease activity is involved in tumor cell invasion (9).
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