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抗体> 凋亡与自噬> Regulation of Apoptosis
产品编号: AC033
产品包装:>20次
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价格: ¥ 1195.34
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产品编号 产品名称 产品包装 产品价格
AC033 Caspase-3(激活型)抗体(兔单抗) >20次 1013.00元
来源 用途 交叉反应性 抗体识别位点 Caspase-3分子量
Rabbit WB, IP, IF, IHC, F H, M, R Cleaved Caspase-3 at Asp175 19/17 kD

WB,Westernblot;IP,Immunoprecipitation;IF,Immunofluorescence;IHC,Immunohistochemistry;F,Flow cytometry.
H,human;M,mouse;R,Rat.
本Caspase-3(激活型)抗体为Cleaved Caspase-3(Asp175)抗体,为进口分装,用人工合成的一段邻近Asp175的human Caspase-3的多肽片段,并通过适当修饰后免疫兔子制备而成的抗Caspase-3(cleaved at Asp175)兔单克隆抗体。克隆号为5A1(Clone 5A1),是一种应用较广的检测Cleaved Caspase-3(Asp175)抗体。
本Caspase-3(激活型)抗体仅仅识别在Asp175位剪切后产生的19和17kDCaspase-3片段,即所谓的激活型Caspase-3。不识别全长的Caspase-3(32kD),也不识别Caspase-3在其它位点被剪切产生的Caspase-3片段。
本抗体可以通过免疫荧光或免疫组化检测细胞凋亡过程中Caspase-3的激活。本抗体染色呈阳性的细胞可以判定为Caspase-3被激活的细胞。
Caspase-3是细胞凋亡过程中最关键的执行分子(key executioner)之一,可以剪切细胞凋亡过程中的许多关键蛋白,例如PARP。Caspase-3的激活需要从没有活性的全长Caspase-3(35kD),在Asp28和Ser29之间及Asp175和Ser176之间进行剪切,产生有活性的17kD亚基和12kD的亚基。
Caspase-3的激活常被作为细胞凋亡的一个重要指标。
配套提供了Western一抗稀释液,可以用于Western检测时的一抗稀释。
建议抗体使用时的稀释比例为(实际使用时需根据抗原水平的高低作适当调整):

WB IP IF IHC F
1:1000 1:100 1:500 1:100 1:100

本抗体如果用于常规的Western检测至少可以检测20次。
包装清单:

产品编号 产品名称 包装
AC033-1 Caspase-3(激活型)抗体(兔单抗) 20μl
AC033-2 Western一抗稀释液 20ml
说明书 1份

保存条件:
Caspase-3(激活型)抗体-20℃保存,Western一抗稀释液-20℃或4℃保存,一年有效。Western一抗稀释液优先推荐4℃保存,长期不使用可以考虑-20℃保存,但冻融可能会导致出现轻微的浑浊和少量不溶物。
注意事项: 
对于本抗体,Western检测时一抗要4℃缓慢摇动过夜,如果仅短时间与一抗孵育检测效果较差。
在Western实验后,请注意回收稀释的抗体。回收的抗体在进行Western实验时至少可以重复使用10次。稀释后的抗体,包括已经使用过的稀释抗体,4℃保存。
回收后重复使用的抗体,使用方法同新鲜稀释的抗体。如果在重复使用过程中发现抗体出现轻微混浊现象,可以10000g离心1-3分钟,取上清用于后续检测。如果回收的抗体出现明显的絮状物或长霉长菌等情况,则可以考虑废弃该抗体。
本产品仅限于专业人员的科学研究用,不得用于临床诊断或治疗,不得用于食品或药品,不得存放于普通住宅内。
为了您的安全和健康,请穿实验服并戴一次性手套操作。

使用说明:
1.Western检测:
A.按照1:1000用碧云天提供的Western一抗稀释液稀释Caspase-3(激活型)抗体。
B.把经过封闭的蛋白膜与稀释好的一抗4℃缓慢摇动过夜,确保稀释的抗体至少能在摇动的瞬间覆盖蛋白膜。
C.回收稀释的一抗,4℃保存以备下次继续使用。
D.按照Western的实验步骤进行后续的洗涤、二抗孵育、洗涤和检测等操作。具体操作可以参考如下网页:http://www.beyotime.com/western.htm
2.免疫染色:
可以使用碧云天生产的免疫染色一抗稀释液(P0103)稀释抗体,使用后注意回收稀释好的一抗,具体操作可以参考如下网页: http://www.beyotime.com/immunol-staining.htm
3.其它实验操作请自行参考适当的protocol进行。
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