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- 详细信息
- 文献和实验
- 技术资料
- 库存:
60
- 英文名:
Capsaicin
- CAS号:
404-86-4
- 供应商:
上海莼试
- 保存条件:
4°C, protect from light
- 规格:
10mM (in 1mL DMSO) 50mg 100mg

规格:10mM (in 1mL DMSO) 50mg 100mg
CAS:404-86-4
别名:(E)-Capsaicin;Zostrix, Qutenza, Axsain, (E)-Capsaicin, Transacin, Capsidol
化学名:(E)-N-[(4-hydroxy-3-methoxyphenyl)methyl]-8-methylnon-6-enamide
Capsaicin分子式:C18H27NO3

分子量:305.41
溶解度:≥ 15.27 mg/mL in DMSO, ≥ 52.3 mg/mL in EtOH with gentle warming
储存条件:4°C, protect from light
General tips:For obtaining a higher solubility , please warm the tube at 37 ℃ and shake it in the ultrasonic bath for a while.
Shipping Condition: Evaluation sample solution : ship with blue ice All other available size: ship with RT , or blue ice upon request
本产品仅供科学实验研究使用! 不能用于临床或动物诊断!
| 产品名称 | Capsaicin | 产品货号 | CS-01Y65452 |
| 规格 | 10mM (in 1mL DMSO) 50mg 100mg | CAS号 | 404-86-4 |
| 含量 | >98.00% | 分子式 | C18H27NO3 |
| 分子量 | 305.41 | 用途 | 仅供科研研究使用 |

Capsaicin is an active component of chili peppers. It selectively binds to TRPV1 which is a heat-activated calcium channel. Capsaicin causes the channel to open below 37 °C. This is why capsaicin is linked to the sensation of heat. [1] Capsaicin has been reported to possess anti-carcinogenic and anti-mutagenic activities. In human glioma A172 cells, capsaicin reduced cell viability with IC50 of ~100 μM treated for 1 day. Capsaicin inhibited cell growth and induced apoptosis through down-regulation of Bcl-2 and activation of caspase-3. Capsaicin also induced terminal differentiation, which contribute to A172 cell growth inhibition.[2] On the other hand, capsaicin reduced the basal generation of ROS, which may played a role in the induction of apoptosis by capsaicin.[3] In MCF-7 cells, Capsaicin induced cell apoptosis through the mitochondrial pathway, and suquently caused PARP-1 cleaved by activation of caspase-7.[4] In human SCLC cell line, capsaicin displayed robust anti-proliferative activity with MTT assay. Furthermore, capsaicin (10 mg/kg body weight) significantly reduced the growth rate of established (800 mm3) H69 tumors xenotransplanted in nude mice. [5]References:1. M. J. Caterina, M. A. Schumacher, M. Tominaga, T. A. Rosen, J. D. Levine and D. Julius, Nature 1997, 389, 816-824.2. Y. G. Gil and M. K. Kang, Life Sci 2008, 82, 997-1003.3. Y. S. Lee, D. H. Nam and J. A. Kim, Cancer Lett 2000, 161, 121-130.4. H. C. Chang, S. T. Chen, S. Y. Chien, S. J. Kuo, H. T. Tsai and D. R. Chen, Hum Exp Toxicol 2011, 30, 1657-1665.5. K. C. Brown, T. R. Witte, W. E. Hardman, H. Luo, Y. C. Chen, A. B. Carpenter, J. K. Lau and P. Dasgupta, PLoS One 2010, 5, e10243. >
使用方法:

1. 常用筛选浓度
注意:用来筛选稳转株的工作浓度需要根据细胞类型,培养基,生长条件和细胞代谢率而变化,推荐使用浓度为50-1000μg/mL。对于第一次使用的实验体系建议通过建立杀灭曲线(kill curve),即剂量反应性曲线,来确定最佳筛选浓度。
一般而言,哺乳动物细胞50-500μg/mL;细菌/植物细胞20-200μg/mL;真菌300-1000μg/mL。
2. 杀灭曲线的建立
注意:为了筛选得到稳定表达目的蛋白的细胞株,需要确定能够杀死未转染宿主细胞的抗生素浓度,可通过建立杀灭曲线(剂量反应曲线)来实现,至少选择5个浓度。
1) 第一天:未转化的细胞按照20-25%的细胞密度铺在合适的培养板上,37℃,CO2培养过夜;注:对于需要更高密度来检测活力的细胞,可增加接种量。
2) 根据细胞类型,设定合适范围内的浓度梯度。以哺乳动物细胞为例,可设定50,100,250,500,750,1000μg/mL。先用去离子水或者PBS buffer按照1:10的比例将母液稀释到5 mg/ml,然后按照下表稀释到相应浓度的工作液。
3) 第二天:替换旧的培养基,换用新鲜配制的含有相应浓度药物的培养基。每个浓度做三个平行孔。
4) 接下来每3-4天更换新的含药物培养基。
5) 按照固定的周期(如每2天)进行活细胞计数来确定阻止未转染细胞生长的恰当浓度。选择在理想的天数(通常是7-10天)内能够杀死绝大多数细胞的浓度为稳定转染细胞筛选用的工作浓度。
3. 稳定转染细胞的筛选
1) 转染48h后,用含有适当浓度的潮霉素B筛选培养基来传代细胞(直接传代或者稀释后传代)。
注意:细胞处于活跃分裂状态时抗生素的杀伤。则当细胞过于稠密,其效率会降低。为了得到较好的筛选效果,最好将细胞稀释至丰度不超过25%
2) 每隔3-4天更换含有药物的筛选培养液。
3) 筛选7天后观察并评估细胞克隆(集落)的形成情况。集落的形成可能还需要一周或者更多的时间,这取决于宿主细胞类型,转染,以及筛选效果。
4) 挑取并转移5-10个抗性克隆于35mm细胞培养板,继续用含药物的筛选培养液维持培养7天。
5) 之后更换正常培养基培养即可。
蛋白酶抑制剂混合物实验步骤:

(1)实验开始前将RNA提取液于65℃水浴锅中预热,离心管中加入ME(巯基乙醇),(10mL加80ul,50mL中加入300ul)
(2)取约0.8g菌丝体(液体培养获得的菌丝用真空抽滤即可!固体培养就更好说了),在液氮中迅速磨成精细粉末,装入50mL离心管,按1g材料8mL的量加入预热的RNA提取液,颠倒混匀
(3)65℃水浴3-10 min,期间混匀2-3次
(4)加入等体积的酚(注意是酸酚pH4.5)::yi戊醇(25:24:1)抽提(10,000rpm,4℃,5 min)
(5)取上清,等体积的yi戊醇(24:1)抽提(10,000rpm,4℃,5 min)
(6)加入1/4V体积10M LiCl溶液,4℃放置6h以上(或过夜)
(7)10,000rpm,4℃离心20min
(8)弃上清,用500ul SSTE溶解沉淀
(9)酚::yi戊醇(25:24:1)抽提两次,:yi戊醇(24:1)抽提1次(10,000rpm,4℃,5min)
(10)加2V体积的无水乙醇,在-70℃冰箱沉淀30min以上
(11)12,000rpm,4℃离心20 min
(12)弃上清.沉淀用70%酒精漂洗一次,干燥
(13)加200ul的DEPC处理水溶解
(14)用非变性琼脂糖凝胶电泳和紫外分光光度计扫描检测RNA的质量(在抽提过程中,若蛋白质含量或其它的杂质还较多,可以增加抽提次数)
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