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- 详细信息
- 文献和实验
- 技术资料
- 库存：
55
- 英文名：
(-)-Huperzine A
- CAS号：
102518-79-6
- 供应商：
上海莼试
- 保存条件：
Store at -20°C
- 规格：
10mM (in 1mL DMSO) 10mg 25mg
本产品仅供科学实验研究使用! 不能用于临床或动物诊断！
化学性质:
(-)-Huperzine A规格：10mM (in 1mL DMSO) 10mg 25mg
CAS：102518-79-6
别名：Huperzine A,
化学名：(5R,9R,E)-5-amino-11-ethylidene-7-methyl-5,6,9,10-tetrahydro-5,9-methanocycloocta[b]pyridin-2(1H)-one
分子式：C15H18N2O
分子量：242.3
溶解度：≥ 12.1mg/mL in DMSO
储存条件：Store at -20°C
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
产品描述:
(?)-Huperzine A (HupA) is an acetylcholinesterase (AChE) inhibitor with an IC50 value of 82 nmol/L [1] and acts as an antagonist of the N-methyl-d-aspartate (NMDA) receptor [2].AChE is the key brain enzyme responsible for the rapid degradation of the neurotransmitter acetylcholine. AChE inhibitors are probably useful in the amelioration of the Alzheimer’s symptomatology [3].It was found that NMDA markedly reduced AChE activities [4]. In rat dissociated hippocampal neurons, HupA inhibited the NMDA-induced current. In neurons, 100 μM HupA, NMDA-induced currents were 55.7 ± 4.9% of the control values. The binding molecular ratio of NMDA receptor: HupA is 1:1. The inhibition of NMDA receptor by HupA is not competitive [5]. HupA significantly increased the phosphorylation levels of both glycogen synthase kinase (GSK)-3α protein and GSK-3β protein in APPsw-overexpressing cells [2]. Activated GSK-3 consequently decreased acetylcholine (ACh) level in the striatum [6].Treated with doses of (?)-huperzine A, AChE?/? mice showed no toxic symptoms and had normal levels of AChE. This demonstrated the specificity of (?)-huperzine A as an inhibitor of AChE at the dose used in vivo [7]. In rat whole brain, oral administration of HupA at a dose of 1.5 μmol/kg (3.6 mg/kg) obtained a maximum inhibition of AChE at 60 min and this maximum inhibition was maintained for 360 min [8].References:[1]. MA Xiao-Chao, XIN Jian, WANG Hai-Xue, et al. Acute effects of huperzine A and tacrine on rat liver. Acta Pharmacol ogica Sinica, 2003, 24(3):247-250.[2]. Zhong Ming Qian and Ya Ke. Huperzine A: is it an effective disease-modifying drug for Alzheimer's disease? Frontiers in Aging Neuroscience, 2014, 6:216.[3]. V. Rajendran, Suo-Bao Rong, Ashima Saxena, et al. Synthesis of a hybrid analog of the acetylcholinesterase inhibitors huperzine A and huperzine B. Tetrahedron Letters, 2001, 42: 5359-5361.[4]. J. R. Delfs, D. M. Saroff, Y. Nishida, et al. Effects of NMDA and its antagonists on ventral horn cholinergic neurons in organotypic roller tube spinal cord cultures. J. Neural Transm., 1997, 104(1):31-51.[5]. J. M. Zhang and G. Y. Hu. Huperzine A, a nootropic alkaloid, inhibits N-methyl-D-aspartate-induced current in rat dissociated hippocampal neurons. Neuroscience, 2001, 105(3):663-9.[6]. L. Zhao, C. B. Chu, J. F. Li, et al. Glycogen synthase kinase-3 reduces acetylcholine level in striatum via disturbing cellular distribution of choline acetyltransferase in cholinergic interneurons in rats. Neuroscience, 2013, 255:203-11.[7]. Ellen G. Duysen, Bin Li, Sultan Darvesh, et al. Sensitivity of butyrylcholinesterase knockout mice to (?)-huperzine A and donepezil suggests humans with butyrylcholinesterase deficiency may not tolerate these Alzheimer’s disease drugs and indicates butyrylcholinesterase function in neurotransmission. Toxicology, 2007, 233:60-69.[8]. Rui Wang, Han Yan and Xi-can Tang. Progress in studies of huperzine A, a natural cholinesterase inhibitor from Chinese herbal medicine. Acta Pharmacologica Sinica, 2006, 27:1-26.
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。
公司正在出售的产品:
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)
| 产品名称 | (-)-Huperzine A | 产品货号 | CS-01Y65053 |
| 规格 | 10mM (in 1mL DMSO) 10mg 25mg | CAS号 | 102518-79-6 |
| 含量 | >98.00% | 分子式 | C15H18N2O |
| 分子量 | 242.3 | 用途 | 仅供科研研究使用 |
(-)-Huperzine A规格：10mM (in 1mL DMSO) 10mg 25mg
CAS：102518-79-6
别名：Huperzine A,
化学名：(5R,9R,E)-5-amino-11-ethylidene-7-methyl-5,6,9,10-tetrahydro-5,9-methanocycloocta[b]pyridin-2(1H)-one
分子式：C15H18N2O
分子量：242.3
溶解度：≥ 12.1mg/mL in DMSO
储存条件：Store at -20°C
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
产品描述:
(?)-Huperzine A (HupA) is an acetylcholinesterase (AChE) inhibitor with an IC50 value of 82 nmol/L [1] and acts as an antagonist of the N-methyl-d-aspartate (NMDA) receptor [2].AChE is the key brain enzyme responsible for the rapid degradation of the neurotransmitter acetylcholine. AChE inhibitors are probably useful in the amelioration of the Alzheimer’s symptomatology [3].It was found that NMDA markedly reduced AChE activities [4]. In rat dissociated hippocampal neurons, HupA inhibited the NMDA-induced current. In neurons, 100 μM HupA, NMDA-induced currents were 55.7 ± 4.9% of the control values. The binding molecular ratio of NMDA receptor: HupA is 1:1. The inhibition of NMDA receptor by HupA is not competitive [5]. HupA significantly increased the phosphorylation levels of both glycogen synthase kinase (GSK)-3α protein and GSK-3β protein in APPsw-overexpressing cells [2]. Activated GSK-3 consequently decreased acetylcholine (ACh) level in the striatum [6].Treated with doses of (?)-huperzine A, AChE?/? mice showed no toxic symptoms and had normal levels of AChE. This demonstrated the specificity of (?)-huperzine A as an inhibitor of AChE at the dose used in vivo [7]. In rat whole brain, oral administration of HupA at a dose of 1.5 μmol/kg (3.6 mg/kg) obtained a maximum inhibition of AChE at 60 min and this maximum inhibition was maintained for 360 min [8].References:[1]. MA Xiao-Chao, XIN Jian, WANG Hai-Xue, et al. Acute effects of huperzine A and tacrine on rat liver. Acta Pharmacol ogica Sinica, 2003, 24(3):247-250.[2]. Zhong Ming Qian and Ya Ke. Huperzine A: is it an effective disease-modifying drug for Alzheimer's disease? Frontiers in Aging Neuroscience, 2014, 6:216.[3]. V. Rajendran, Suo-Bao Rong, Ashima Saxena, et al. Synthesis of a hybrid analog of the acetylcholinesterase inhibitors huperzine A and huperzine B. Tetrahedron Letters, 2001, 42: 5359-5361.[4]. J. R. Delfs, D. M. Saroff, Y. Nishida, et al. Effects of NMDA and its antagonists on ventral horn cholinergic neurons in organotypic roller tube spinal cord cultures. J. Neural Transm., 1997, 104(1):31-51.[5]. J. M. Zhang and G. Y. Hu. Huperzine A, a nootropic alkaloid, inhibits N-methyl-D-aspartate-induced current in rat dissociated hippocampal neurons. Neuroscience, 2001, 105(3):663-9.[6]. L. Zhao, C. B. Chu, J. F. Li, et al. Glycogen synthase kinase-3 reduces acetylcholine level in striatum via disturbing cellular distribution of choline acetyltransferase in cholinergic interneurons in rats. Neuroscience, 2013, 255:203-11.[7]. Ellen G. Duysen, Bin Li, Sultan Darvesh, et al. Sensitivity of butyrylcholinesterase knockout mice to (?)-huperzine A and donepezil suggests humans with butyrylcholinesterase deficiency may not tolerate these Alzheimer’s disease drugs and indicates butyrylcholinesterase function in neurotransmission. Toxicology, 2007, 233:60-69.[8]. Rui Wang, Han Yan and Xi-can Tang. Progress in studies of huperzine A, a natural cholinesterase inhibitor from Chinese herbal medicine. Acta Pharmacologica Sinica, 2006, 27:1-26.
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。
公司正在出售的产品:
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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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文献和实验相关实验
3周内可恢复。某些病例随剂量减少，ALT也可恢复正常。tacrine引起ALT升高的机制目前尚不清楚，一般认为是本品的代谢产物介导的毒性所致。其他不良反应包括尿频、流涎、多汗、眩晕和皮疹等。 石杉碱甲 Huperzine A Huperzine A也称哈伯因、双益平等，是中国学者从天然植物中提取的一种生物碱，是一种高选择性胆碱酯酶抑制剂，90年代初被卫生部批准为治疗早老性痴呆症的新药。石杉碱甲具有显著的改善记忆和认知功能的作用，药理与临床研究均表明，它明显优于国外
amyloid peptide[J]. Brain Res,2001,912(2):128-36. [6] Wang R , Zhang HY, Tang XC. Huperzine A attenuates cognitive dysfunction and neuronal degeneration caused byβ-amyloid protein-(1～40) in rat[J]. Eur J Pharmacol,2001,421(3):149-56. [7] Yamaguchi Y
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(-)-Huperzine A
￥600 - 3200
