Cell Meter 荧光法胞内总ROS检测试剂盒 适合于流式细胞仪

货号22904存储条件f/l
规格100 Tests价格3720
Ex (nm)490Em (nm)520
分子量溶剂
产品详细介绍


简要概述

活性氧(ROS)是氧正常代谢的天然副产物,在细胞信号传导中起重要作用。但是,在与氧化应激相关的状态下,ROS水平会急剧增加。 ROS的积累会严重破坏细胞结构。氧化应激在心血管疾病,糖尿病,骨质疏松症,中风,炎性疾病,许多神经退行性疾病和癌症中的作用已得到公认。 ROS检测将有助于确定氧化应激如何调节各种细胞内途径。 Cell Meter 荧光法胞内总ROS检测试剂盒使用我们独特的Amplite ROS Green荧光定量活细胞中的ROS,Amplite ROS Green具有细胞渗透性。与ROS反应时会生成绿色荧光。 Cell Meter 荧光法胞内总ROS检测试剂盒提供了一种灵敏的一步荧光测定法,可在培养一小时后检测活细胞中的细胞内ROS。该试剂盒针对流式细胞仪应用进行了优化,其信号可以通过Ex / Em = 490/520 nm(FL1通道)进行检测。百萤生物是AAT Bioquest的中国代理商,为您提供最优质的Cell Meter 荧光法胞内总ROS检测试剂盒。 



产品说明书

样品实验方案

简要概述

1.准备密度为0.5-1×106细胞/ mL的细胞
2.在0.5 mL细胞悬液中加入1 µL 500X Amplite ROS Green
3.在37℃下将细胞染色1小时
4.处理细胞以诱导ROS
5.使用带有FL1通道的流式细胞仪分析细胞(Ex / Em = 490/520 nm)

 

溶液配制

1.储备溶液配制

        除非另有说明,否则所有未使用的储备溶液应分成一次性等分试样,并在制备后储存在-20°C。 避免反复冻融循环。

Amplite ROS绿色储备溶液(500X):将100 µL DMSO(组分C)添加到Amplite ROS Green(组分A)的小瓶中,并充分混合以制成500X Amplite ROS Green储备液。 避光。 注意:要存放,请密封管。有关细胞样品制备的指南,请点击查看

 

操作步骤

1.对于每个样品,以0.5×105至1×106细胞/ mL的密度在0.5 mL测定缓冲液(组分B)或自备缓冲液中制备细胞。注意:应单独评估每个细胞系,以确定诱导ROS的最佳细胞密度。

2.将1 µL 500X Amplite ROS Green储备溶液加入0.5 mL细胞悬液中。

3.在37ºC下孵育1小时。注意:对于贴壁细胞,用0.5 mM EDTA轻轻提起细胞以保持细胞完整,并在与Amplite ROS Green孵育之前用含血清的培养基洗涤细胞一次。合适的孵育时间取决于所用的单个细胞类型和测试化合物,优化每个实验的孵育时间。

4.通过在所需的缓冲液(例如PBS或HBSS)中添加50 µL 11X测试化合物来处理细胞。对于对照孔(未处的细胞),添加相应量的缓冲液。

5.将细胞在37ºC下孵育,以诱导ROS(避光)。注意:我们在37℃下用100 µM TBHP(氢过氧化叔丁基)处理Jurkat细胞30分钟,以诱导ROS。有关详细信息,请参见图1。

6.使用具有FL1通道(Ex / Em = 490/520 nm)的流式细胞仪检测荧光强度。

 

图示

图1.使用Cell Meter 荧光法细胞内总ROS活性测定试剂盒在TBHP处理后检测Jurkat细胞中的细胞内ROS。 将细胞与Amplite ROS Green在37°C孵育1小时。 然后将细胞在不使用(蓝色)或(红色)100 µM TBHP的情况下在37°C下处理30分钟。 使用流式细胞仪(BD FACSCalibur)在FL1通道处检测荧光信号。

 

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Notoginsenoside R1 attenuates high glucose-induced endothelial damage in rat retinal capillary endothelial cells by modulating the intracellular redox state
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Good hydration and cell-biological performances of superparamagnetic calcium phosphate cement with concentration-dependent osteogenesis and angiogenesis induced by ferric iron
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Topiramate Protects Pericytes from Glucotoxicity: Role for Mitochondrial CA VA in Cerebromicrovascular Disease in Diabetes
Authors: Patrick, Ping and Price, Tulin O and Diogo, Ana L and Sheibani, Nader and Banks, William A and Shah, Gul N
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Down-regulated peroxisome proliferator-activated receptor γ (PPARγ) in lung epithelial cells promotes a PPARγ agonist-reversible proinflammatory phenotype in chronic obstructive pulmonary disease (COPD)
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Xanthine oxidase inhibition by febuxostat attenuates experimental atherosclerosis in mice
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说明书
Cell Meter 荧光法胞内总ROS检测试剂盒 适合于流式细胞仪 .pdf