Product Name | HCV NS3/4A protease genotype 1b, recombinant |
Size | 5 µg |
Catalog # | AS-61017-5 |
US$ | $247 |
The NS3 protease of hepatitis C virus (HCV) is responsible for the cleavage at the NS3/NS4A, NS4A/NS4B, NS4B/NS5A, and NS5A/NS5B sites of the nonstructural protein. It is essential for viral replication and the formation of infectious viral particles, and thus has been considered as one of the most attractive targets for anti-HCV therapy. Recombinant HCV NS3/4A protease is highly active and can be used with SensoLyte® 490 or 520 HCV Protease Assay Kits (Cat# 71126, 71145) to screen for anti-HCV protease drugs. | |
Detailed Information | DatasheetMaterial Safety Data Sheets (MSDS)Poster |
Storage | Store at -80°C. Avoid multiple thaw-freeze cycles. |
References | 1. R. Bartenschlager, R. et al. J.Virol. 67, 3835-3844 (1993).2. Eckart, MR. et al. Biochem.Biophys.Res.Commun. 192, 399-406 (1993).3. Landro, JA. et al. Biochemistry 36, 9340-9348 (1997).4. Kim, JL. et al. Cell 87, 343-355 (1996).5. Love, RA. et al. Cell 87, 331-342 (1996). |
Product Citations | Ma, C. et al. (2009). HCV Protease Inhibitory, Cytotoxic and Apoptosis-Inducing Effects of Oleanolic Acid Derivatives. J Pharm Pharmaceut Sci 12, 243.Phuong, DT. et al. (2009). Inhibitory effects of antrodins A-E from Antrodia cinnamomea and their metabolites on hepatitis C virus protease. Phytother Res 23, 582.of antrodins A-E from Antrodia cinnamomea and their metabolites on hepatitis C virus protease. Phytother. Res. 23, 582.Yu, X. (2009). Development of a cell-based hepatitis C virus (HCV) infection FRET assay for high throughput antiviral compound screening. Antimicrob Agents Chemo doi:10.1128/AAC.00495-09.Ma, C. et al. (2007). Triterpenes from Cynomorium songariciumanalysis of HCV protease inhibitory activity, quantification, and content change under the influence of heating. J Nat Med 63, 9. |
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1.将外源基因插入慢病毒载体
2.将构建完成的载体与慢病毒包装质粒混合,共转染靶细胞
3.收集病毒液
4.用病毒液感染靶细胞
5.用载体上带的抗生素进行筛选,如果没有,可以用无限稀释法
6.获得稳转株
对于瞬时转染的检测:如果有报告基因,就通过报告基因的表达看转染成功与否;没有报告基因的话,就在24~96h内收获细胞通过RT-PCR和WB来鉴定。
推荐了解“主细胞库”“工作细胞库”这两个名词。
① 在构建载体时,目的基因直接整合到细胞染色体组上,最好不要通过先瞬转在筛选稳定细胞株的这种方法,因为转染效率没有保证
② 高表达载体的构建,哺乳动物表达量一直是它自身的缺点,最好根据高表达载体定向的驯化细胞,提高蛋白表达量
③ 细胞的选择,筛选稳定细胞株我们常用的细胞是CHO,中国仓鼠卵巢细胞,由于CHO具有诸多的优点因此适合用于筛选稳定细胞株,而HEK293细胞则常用于瞬时转染
④ 后期的筛选,双抗预防污染,筛选细胞的时候抗生素浓度一定要做预实验,而且转染的时候不能有抗生素,关于细胞转染 稳定细胞系构建的相关理论
至于到底好不好转就跟你的细胞有关了,和细胞是否是稳转株关系不大。
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