Screen Quest Calbryte 590 免丙磺舒和免洗钙检测试剂盒 货号36202-AAT Bioquest荧光染料

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Screen Quest Calbryte 590 免丙磺舒和免洗钙检测试剂盒

Screen Quest Calbryte 590 免丙磺舒和免洗钙检测试剂盒

货号 36202 存储条件 在零下15度以下保存, 避免光照
规格 100 Plates 价格 92016
Ex (nm) 581 Em (nm) 593
分子量 溶剂
产品详细介绍

简要概述

Screen Quest Calbryte 590 免丙磺舒和免洗钙检测试剂盒是美国AAT Bioquest研发的用于检测钙离子的试剂盒钙通量测定是用于筛选G蛋白偶联受体(GPCR)的药物发现中的优选方法。 Screen Quest Calbryte-590不含Probenecid和Wash-Free的钙测定试剂盒提供最强大的均相荧光测定,用于检测细胞内钙动员。表达感兴趣的GPCR的细胞通过钙预先加载我们专有的Calbryte -590NW,其可以穿过细胞膜。 Calbryte -590 NW是最适合HTS筛查的钙指示剂。一旦进入细胞内,Calbryte -590NW的亲脂性阻断基团被非特异性细胞酯酶切割,导致带电荷的荧光染料停留在细胞内,并且在与钙结合后其荧光大大增强。当用筛选化合物刺激细胞时,该受体表明细胞内钙的释放,这极大地增加了Calbryte -590NW的荧光。其优异的细胞保留特性,高灵敏度和100-250倍的荧光增加(当它与钙形成复合物时)使Calbryte -590NW成为测量细胞钙的理想指标.Calbryte -590NW是唯一的钙染料不需要丙磺舒以获得更好的细胞保留。这款Screen Quest Calbryte-590不含Probenecid和Wash-Free的钙测定试剂盒提供了最优化的检测方法,用于监测G蛋白偶联受体(GPCR)和钙通道与脆弱或困难的细胞系。该测定可以以方便的96孔或384孔微量滴定板形式进行,并且易于适应自动化。金畔生物是AAT Bioquest 的中国代理商,为您提供最优质的钙检测试剂盒。

 

适用仪器


荧光酶标仪  
激发: 540nm
发射: 590nm
cutoff: 570nm
推荐孔板: 黑色透明
读取模式: 底读模式
其他仪器
FDSS, ViewLux, NOVOStar, ArrayScan, FlexStation, IN Cell Analyzer

产品说明书

样品实验方案

简要概述

  1. 在生长培养基中准备细胞
  2. 添加Calbryte 590 AM染料加载溶液(对于96孔板为100 µL /孔,对于384孔板为25 µL /孔)
  3. 在室温或37°C下孵育45-60分钟
  4. 在Ex / Em = 540/590 nm处检测荧光

 

溶液配制

储备溶液配制

1. Calbryte 590 AM储备溶液:向Calbryte 590 AM(组分A)的小瓶中加入20 µL(#36200)或200 µL(#36201和#36202)DMSO。 注意:20 µL Calbryte 590 AM储备溶液足以装一板。 未用完的Calbryte 590 AM储备溶液可以等分分装,并在<-20℃下保存。 注意:避光,并避免重复的冻融循环。

2.分析缓冲液(1X):将9 mL的HHBS(组分C,试剂盒#36202中未包括)与1 mL的10XPluronic®F127 Plus(10X)(组分B)充分混合。

 

工作溶液配制

Calbryte 590 AM工作溶液:将20 µL Calbryte 590 AM储备溶液添加到10 mL的测定缓冲液(1X)中,并充分混合。 注意:该工作溶液在室温下至少可稳定2小时。 注意:10 mL染料加载溶液足以用于一块96孔板。

 

实验步骤

1.将100 µL /孔(96孔板)或25 µL /孔(384孔板)的Calbryte 590 AM染料加载溶液添加到细胞板中。

2.在细胞培养箱中将染料加载板孵育60分钟,然后在室温下将板再孵育15-30分钟。 注意:如果测定需要37°C,请立即进行实验,而无需进一步室温孵育。 如果细胞在室温下能长时间正常工作,则在室温下孵育细胞板1小时(建议孵育时间不超过2小时)。

3.用HHBS或所需的缓冲液准备复合板。

4.通过检测Ex / Em = 540/590 nm的荧光强度。

 

图示

Screen Quest  Calbryte 590 免丙磺舒和免洗钙检测试剂盒    货号36202

图1.图表说明了信噪比(SNR)x 100%。 使用Screen Quest Calbryte 590无丙磺舒和无洗涤钙测定试剂盒测量CHO-K1细胞中的ATP剂量反应。 将CHO-K1细胞以50,000个细胞/ 100 µL /孔在96孔黑色板上接种过夜。 加入100 µL的染料加载溶液,在37°C下孵育45分钟,在室温下孵育15分钟。 FlexStation 3添加了ATP(50 µL /孔)以达到最终指示的浓度。

 

 

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说明书
Screen Quest Calbryte 590 免丙磺舒和免洗钙检测试剂盒 .pdf