赛信通SignalSilence® MTAP siRNA I可以通过RNA干扰技术特异性地抑制MTAP的表达(RNA干扰技术是通过往细胞内导入双链干扰RNA来选择性地抑制某些基因的表达)。赛信通所有的SignalSilence® siRNA产品都经过了实验室的反复验证,并且通过Western实验来证实了基因表达的抑制效果。寡核苷酸的合成由三苯甲基分析逐一检测碱基序列,以确保合适的耦合效率,然后通过亲和固相萃取纯化寡核苷酸。退火的双链RNA进一步通过质谱分析,以验证双链的确切组成。每批次产品利用质谱检测,以最大程度地确保批次间的一致性。CST with any questions on use.CST推荐对细胞用100 nM MTAP siRNA I转染48到72小时后再进行裂解。转染步骤参考转染试剂的厂家产品说明。如果在使用过程中有任何问题,请联系CST技术支持。MTAP是腺膘呤和蛋氨酸回收反应中的必需酶。MTAP可以催化5''-甲基硫腺苷酸的裂解,生成腺苷酸和5-甲硫基-D-核糖-1磷酸。腺苷酸然后用于合成AMP,而5-甲硫基-D-核糖-1磷酸则转化成蛋氨酸(1,2)。MTAP在大多数正常组织和细胞中都有表达,但在多种人肿瘤细胞中缺失,包括前列腺腺瘤,神经内分泌肿瘤,非小细胞肺癌和乳腺癌。MTAP经常与p16 (cdkN2a/ARF)共同缺失(3-5)。在琼脂糖凝胶上,对乳腺细胞过表达MTAP会抑制其克隆形成能力,因此可能预示着它有着肿瘤抑制效应(6)。
SignalSilence® MTAP siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit MTAP 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 MTAP 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
Background
MTAP is an enzyme that is essential for the salvage pathway for both adenine and methionine synthesis. MTAP catalyzes the cleavage of 5’-methylthioadenosine into adenine and 5-methylthio-D-ribose-1-phosphate. Adenine is then used to generate AMP whereas 5-methylthio-D-ribose-1-phosphate is converted into methionine (1,2). MTAP is expressed in all normal cells and tissues, although frequently lost in different human tumors including pancreatic adenocarcinoma, neuroendocrine tumors, non-small cell lung carcinoma and breast carcinoma. MTAP is usually codeleted with p16 (cdkN2a/ARF) (3-5). MTAP overexpression in breast cancer cells inhibits their ability to form colonies in soft agar, thereby implicating its function as a tumor suppressor (6).