Screen Quest Calbryte 590 免丙磺舒和免洗钙检测试剂盒 货号36200-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 36200 存储条件 在零下15度以下保存, 避免光照
规格 1 Plate 价格 3840
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36200

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