PhosphoWorks 发光法ATP检测试剂盒* DTT – Free* 货号21612-AAT Bioquest荧光染料

上海金畔生物科技有限公司代理AAT Bioquest荧光染料全线产品,欢迎访问AAT Bioquest荧光染料官网了解更多信息。

PhosphoWorks 发光法ATP检测试剂盒* DTT – Free*

PhosphoWorks 发光法ATP检测试剂盒* DTT – Free*

PhosphoWorks 发光法ATP检测试剂盒* DTT - Free*    货号21612 货号 21612 存储条件 在零下15度以下保存, 避免光照
规格 1 Plate 价格 1944
Ex (nm) Em (nm)
分子量 溶剂
产品详细介绍

简要概述

PhosphoWorks 发光法ATP检测试剂盒是美国AAT Bioquest生产的用于检测ATP的试剂盒,腺苷三磷酸(ATP)在细胞能量学,代谢调节和细胞信号传导中起重要作用。 PhosphoWorks ATP检测试剂盒提供快速,简单和均匀的发光检测,用于测定哺乳动物细胞中的细胞增殖和细胞毒性。 可以以方便的96孔和384孔微量滴定板形式进行测定。 该测定的高灵敏度允许在许多生物系统,环境样品和食物中检测ATP。 该PhosphoWorks ATP检测试剂盒不使用DTT,并且具有长达4小时的稳定发光信号。 它具有稳定的发光,不需要混合或分离,并配制成具有最小的手动操作时间。金畔生物是AAT Bioquest的中国代理商,为您提供最优质的PhosphoWorks 发光法ATP检测试剂盒。 

 

适用仪器


发光酶标仪  
推荐孔板: 白色孔板

产品说明书

操作步骤

简要概述

1.用测试化合物(100μL/ 96孔板或25μL/ 384孔板)制备细胞(样品)
2.加入等体积的ATP工作溶液(100μL/ 96孔板或25μL/ 384孔板)
3.在室温下孵育10-20分钟
4.监测发光强度

 

制备工作溶液

1.将10 mL反应缓冲液(组分C)转移到ATP传感器(组分B)中并充分混合。

2.将20μLATP监测酶(组分A)加入到组分B + C的瓶中并充分混合以制备ATP工作溶液。 注意:避免来自外源生物来源的潜在ATP污染。

有关细胞样品制备的指南(点击查看)

 

样品实验方案

1.运行ATP测定:

1.1用测试化合物处理细胞(或样品),在96孔板中加入10μL10X化合物,或在所需化合物缓冲液中加入5μL5X化合物用于384孔板。对于空白孔(没有细胞的培养基),加入相应量的化合物缓冲液。

1.2将细胞板在37℃,5%CO 2培养箱中孵育所需的一段时间,例如24,48或96小时。

1.3向每个孔中加入100μL(96孔板)或25μL(384孔板)的ATP工作溶液。

1.4在室温下孵育10-20分钟。

1.5用标准发光计监测发光强度。

 

2.生成标准ATP校准曲线:

        如果需要计算样品中ATP的绝对量,则应与上述分析一起生成ATP标准曲线。

2.1通过包含不含ATP的样品(作为对照)在含有0.1%BSA的PBS缓冲液中制备ATP系列稀释液以测量背景发光。注意:通常ATP浓度范围为0.1 nM至1μM是合适的。

2.2将相同量的稀释的ATP溶液加入空板中(对于96孔板为100μL或对于384孔板为25μL)。

2.3加入100μL/孔(96孔板)或25μL/孔(384孔板)的ATP工作溶液。

2.4将反应混合物在室温下孵育10至20分钟。

2.5用标准发光计记录发光强度。

2.6生成ATP标准曲线。

 

数据分析

        从空白标准孔获得的读数(RLU)用作阴性对照。 从其他标准的读数中减去该值,以获得基线校正值。 然后,绘制标准读数以获得标准曲线和方程。 该等式可用于计算ATP样品。 我们建议使用在线线性回归计算器,该计算器可在以下位置找到:

PhosphoWorks 发光法ATP检测试剂盒* DTT - Free*    货号21612

图1.使用NOVOstar板读数器(BMG Labtech)在96孔白板上用PhosphoWorks TM发光ATP测定试剂盒* DTT-Free *测量ATP剂量响应。 该试剂盒可在20分钟内检测到(3 pmole /孔)0.03 nM ATP。 积分时间为1秒。 半衰期超过2小时。

 

参考文献

High throughput cell-based assay for identification of glycolate oxidase inhibitors as a potential treatment for Primary Hyperoxaluria Type 1
Authors: Mengqiao Wang, Miao Xu, Yan Long, Sonia Fargue, Noel Southall, Xin Hu, John C McKew, Christopher J Danpure, Wei Zheng
Journal: Scientific Reports (2016)

NT1014, a novel biguanide, inhibits ovarian cancer growth in vitro and in vivo
Authors: Lu Zhang, Jianjun Han, Amanda L Jackson, Leslie N Clark, Joshua Kilgore, Hui Guo, Nick Livingston, Kenneth Batchelor, Yajie Yin, Timothy P Gilliam
Journal: Journal of Hematology & Oncology (2016): 91

The Different Effects of Atorvastatin and Pravastatin on Cell Death and PARP Activity in Pancreatic NIT-1 Cells
Authors: Ya-Hui Chen, Yi-Chun Chen, Chin-San Liu, Ming-Chia Hsieh
Journal: Journal of Diabetes Research (2016)

BPA-induced DNA hypermethylation of the master mitochondrial gene PGC-1α contributes to cardiomyopathy in male rats
Authors: Ying Jiang, Wei Xia, Jie Yang, Yingshuang Zhu, Huailong Chang, Juan Liu, Wenqian Huo, Bing Xu, Xi Chen, Yuanyuan Li
Journal: Toxicology (2015): 21–31

Glutamine promotes ovarian cancer cell proliferation through the mTOR/S6 pathway
Authors: Lingqin Yuan, Xiugui Sheng, Adam K Willson, Dario R Roque, Jessica E Stine, Hui Guo, Hannah M Jones, Chunxiao Zhou, Victoria L Bae-Jump
Journal: Endocrine-related cancer (2015): 577–591

JQ1 suppresses tumor growth through downregulating LDHA in ovarian cancer
Authors: Haifeng Qiu, Amanda L Jackson, Joshua E Kilgore, Yan Zhong, Leo Li-Ying Chan, Paola A Gehrig, Chunxiao Zhou, Victoria L Bae-Jump
Journal: Oncotarget (2015): 6915

Loss of histone deacetylase Hdac1 disrupts metabolic processes in intestinal epithelial cells
Authors: Alexis Gonneaud, Naomie Turgeon, Frančois-Michel Boisvert, Frančois Boudreau, Claude Asselin
Journal: FEBS letters (2015): 2776–2783

A neutrophil intrinsic impairment affecting Rab27a and degranulation in cystic fibrosis is corrected by CFTR potentiator therapy
Authors: Kerstin Pohl, Elaine Hayes, Joanne Keenan, Michael Henry, Paula Meleady, Kevin Molloy, Bakr Jundi, David A Bergin, Cormac McCarthy, Oliver J McElvaney
Journal: Blood (2014): 999–1009

Fluorescence lifetime imaging unravels C. trachomatis metabolism and its crosstalk with the host cell
Authors: Márta Szaszák, Philipp Steven, Kensuke Shima, Regina Orzekowsky-Schroder, Gereon Huttmann, Inke R Konig, Werner Solbach, Jan Rupp
Journal: PLoS pathogens (2011): e1002108

G protein coupled receptor kinase 2 interacting protein 1 (GIT1) is a novel regulator of mitochondrial biogenesis in heart
Authors: Jinjiang Pang, Xiangbin Xu, Michael R Getman, Xi Shi, Stephen L Belmonte, Heidi Michaloski, Amy Mohan, Burns C Blaxall, Bradford C Berk
Journal: Journal of molecular and cellular cardiology (2011): 769–776

 

相关产品

产品名称 货号
PhosphoWorks 发光法ATP检测试剂盒 *温和发光* Cat#21609
PhosphoWorks 发光法ATP检测试剂盒 *明亮发光* Cat#21610
PhosphoWorks 荧光ATP检测试剂盒 Cat#21620

说明书
PhosphoWorks 发光法ATP检测试剂盒* DTT – Free*.pdf

PhosphoWorks 发光法ATP检测试剂盒* DTT – Free* 货号21612-AAT Bioquest荧光染料

上海金畔生物科技有限公司代理AAT Bioquest荧光染料全线产品,欢迎访问AAT Bioquest荧光染料官网了解更多信息。

PhosphoWorks 发光法ATP检测试剂盒* DTT – Free*

PhosphoWorks 发光法ATP检测试剂盒* DTT – Free*

PhosphoWorks 发光法ATP检测试剂盒* DTT - Free*    货号21612 货号 21612 存储条件 在零下15度以下保存, 避免光照
规格 1 Plate 价格 1944
Ex (nm) Em (nm)
分子量 溶剂
产品详细介绍

简要概述

PhosphoWorks 发光法ATP检测试剂盒是美国AAT Bioquest生产的用于检测ATP的试剂盒,腺苷三磷酸(ATP)在细胞能量学,代谢调节和细胞信号传导中起重要作用。 PhosphoWorks ATP检测试剂盒提供快速,简单和均匀的发光检测,用于测定哺乳动物细胞中的细胞增殖和细胞毒性。 可以以方便的96孔和384孔微量滴定板形式进行测定。 该测定的高灵敏度允许在许多生物系统,环境样品和食物中检测ATP。 该PhosphoWorks ATP检测试剂盒不使用DTT,并且具有长达4小时的稳定发光信号。 它具有稳定的发光,不需要混合或分离,并配制成具有最小的手动操作时间。金畔生物是AAT Bioquest的中国代理商,为您提供最优质的PhosphoWorks 发光法ATP检测试剂盒。 

 

适用仪器


发光酶标仪  
推荐孔板: 白色孔板

产品说明书

操作步骤

简要概述

1.用测试化合物(100μL/ 96孔板或25μL/ 384孔板)制备细胞(样品)
2.加入等体积的ATP工作溶液(100μL/ 96孔板或25μL/ 384孔板)
3.在室温下孵育10-20分钟
4.监测发光强度

 

制备工作溶液

1.将10 mL反应缓冲液(组分C)转移到ATP传感器(组分B)中并充分混合。

2.将20μLATP监测酶(组分A)加入到组分B + C的瓶中并充分混合以制备ATP工作溶液。 注意:避免来自外源生物来源的潜在ATP污染。

有关细胞样品制备的指南(点击查看)

 

样品实验方案

1.运行ATP测定:

1.1用测试化合物处理细胞(或样品),在96孔板中加入10μL10X化合物,或在所需化合物缓冲液中加入5μL5X化合物用于384孔板。对于空白孔(没有细胞的培养基),加入相应量的化合物缓冲液。

1.2将细胞板在37℃,5%CO 2培养箱中孵育所需的一段时间,例如24,48或96小时。

1.3向每个孔中加入100μL(96孔板)或25μL(384孔板)的ATP工作溶液。

1.4在室温下孵育10-20分钟。

1.5用标准发光计监测发光强度。

 

2.生成标准ATP校准曲线:

        如果需要计算样品中ATP的绝对量,则应与上述分析一起生成ATP标准曲线。

2.1通过包含不含ATP的样品(作为对照)在含有0.1%BSA的PBS缓冲液中制备ATP系列稀释液以测量背景发光。注意:通常ATP浓度范围为0.1 nM至1μM是合适的。

2.2将相同量的稀释的ATP溶液加入空板中(对于96孔板为100μL或对于384孔板为25μL)。

2.3加入100μL/孔(96孔板)或25μL/孔(384孔板)的ATP工作溶液。

2.4将反应混合物在室温下孵育10至20分钟。

2.5用标准发光计记录发光强度。

2.6生成ATP标准曲线。

 

数据分析

        从空白标准孔获得的读数(RLU)用作阴性对照。 从其他标准的读数中减去该值,以获得基线校正值。 然后,绘制标准读数以获得标准曲线和方程。 该等式可用于计算ATP样品。 我们建议使用在线线性回归计算器,该计算器可在以下位置找到:

PhosphoWorks 发光法ATP检测试剂盒* DTT - Free*    货号21612

图1.使用NOVOstar板读数器(BMG Labtech)在96孔白板上用PhosphoWorks TM发光ATP测定试剂盒* DTT-Free *测量ATP剂量响应。 该试剂盒可在20分钟内检测到(3 pmole /孔)0.03 nM ATP。 积分时间为1秒。 半衰期超过2小时。

 

参考文献

High throughput cell-based assay for identification of glycolate oxidase inhibitors as a potential treatment for Primary Hyperoxaluria Type 1
Authors: Mengqiao Wang, Miao Xu, Yan Long, Sonia Fargue, Noel Southall, Xin Hu, John C McKew, Christopher J Danpure, Wei Zheng
Journal: Scientific Reports (2016)

NT1014, a novel biguanide, inhibits ovarian cancer growth in vitro and in vivo
Authors: Lu Zhang, Jianjun Han, Amanda L Jackson, Leslie N Clark, Joshua Kilgore, Hui Guo, Nick Livingston, Kenneth Batchelor, Yajie Yin, Timothy P Gilliam
Journal: Journal of Hematology & Oncology (2016): 91

The Different Effects of Atorvastatin and Pravastatin on Cell Death and PARP Activity in Pancreatic NIT-1 Cells
Authors: Ya-Hui Chen, Yi-Chun Chen, Chin-San Liu, Ming-Chia Hsieh
Journal: Journal of Diabetes Research (2016)

BPA-induced DNA hypermethylation of the master mitochondrial gene PGC-1α contributes to cardiomyopathy in male rats
Authors: Ying Jiang, Wei Xia, Jie Yang, Yingshuang Zhu, Huailong Chang, Juan Liu, Wenqian Huo, Bing Xu, Xi Chen, Yuanyuan Li
Journal: Toxicology (2015): 21–31

Glutamine promotes ovarian cancer cell proliferation through the mTOR/S6 pathway
Authors: Lingqin Yuan, Xiugui Sheng, Adam K Willson, Dario R Roque, Jessica E Stine, Hui Guo, Hannah M Jones, Chunxiao Zhou, Victoria L Bae-Jump
Journal: Endocrine-related cancer (2015): 577–591

JQ1 suppresses tumor growth through downregulating LDHA in ovarian cancer
Authors: Haifeng Qiu, Amanda L Jackson, Joshua E Kilgore, Yan Zhong, Leo Li-Ying Chan, Paola A Gehrig, Chunxiao Zhou, Victoria L Bae-Jump
Journal: Oncotarget (2015): 6915

Loss of histone deacetylase Hdac1 disrupts metabolic processes in intestinal epithelial cells
Authors: Alexis Gonneaud, Naomie Turgeon, Frančois-Michel Boisvert, Frančois Boudreau, Claude Asselin
Journal: FEBS letters (2015): 2776–2783

A neutrophil intrinsic impairment affecting Rab27a and degranulation in cystic fibrosis is corrected by CFTR potentiator therapy
Authors: Kerstin Pohl, Elaine Hayes, Joanne Keenan, Michael Henry, Paula Meleady, Kevin Molloy, Bakr Jundi, David A Bergin, Cormac McCarthy, Oliver J McElvaney
Journal: Blood (2014): 999–1009

Fluorescence lifetime imaging unravels C. trachomatis metabolism and its crosstalk with the host cell
Authors: Márta Szaszák, Philipp Steven, Kensuke Shima, Regina Orzekowsky-Schroder, Gereon Huttmann, Inke R Konig, Werner Solbach, Jan Rupp
Journal: PLoS pathogens (2011): e1002108

G protein coupled receptor kinase 2 interacting protein 1 (GIT1) is a novel regulator of mitochondrial biogenesis in heart
Authors: Jinjiang Pang, Xiangbin Xu, Michael R Getman, Xi Shi, Stephen L Belmonte, Heidi Michaloski, Amy Mohan, Burns C Blaxall, Bradford C Berk
Journal: Journal of molecular and cellular cardiology (2011): 769–776

 

相关产品

产品名称 货号
PhosphoWorks 发光法ATP检测试剂盒 *温和发光* Cat#21609
PhosphoWorks 发光法ATP检测试剂盒 *明亮发光* Cat#21610
PhosphoWorks 荧光ATP检测试剂盒 Cat#21620

说明书
PhosphoWorks 发光法ATP检测试剂盒* DTT – Free*.pdf

PhosphoWorks 发光法ATP检测试剂盒* DTT – Free* 货号21613-AAT Bioquest荧光染料

上海金畔生物科技有限公司代理AAT Bioquest荧光染料全线产品,欢迎访问AAT Bioquest荧光染料官网了解更多信息。

PhosphoWorks 发光法ATP检测试剂盒* DTT – Free*

PhosphoWorks 发光法ATP检测试剂盒* DTT – Free*

PhosphoWorks 发光法ATP检测试剂盒* DTT - Free*    货号21613 货号 21613 存储条件 在零下15度以下保存, 避免光照
规格 10 Plates 价格 12588
Ex (nm) Em (nm)
分子量 溶剂
产品详细介绍

简要概述

PhosphoWorks 发光法ATP检测试剂盒* DTT – Free*是美国AAT Bioquest生产的用于检测ATP的试剂盒,三磷酸腺苷(ATP)在细胞能量生成,代谢调节和细胞信号传导中起着基本作用。 PhosphoWorks ATP检测试剂盒提供了一种快速,简单且均一的发光检测方法,用于测定哺乳动物细胞中的细胞增殖和细胞毒性。 该测定可以以方便的96孔和384孔微量滴定板形式进行。 此测定法的高灵敏度允许在许多生物系统,环境样品和食品中检测ATP。 此PhosphoWorks ATP检测试剂盒不使用DTT,并且具有长达4小时的稳定发光信号。 它具有稳定的发光,无需混合或分离,并且配方具有最小的动手时间。金畔生物是AAT Bioquest的中国代理商,为您提供最优质的PhosphoWorks 发光法ATP检测试剂盒* DTT – Free*。 

 

适用仪器


发光酶标仪  
推荐孔板: 白色孔板

产品说明书

操作步骤

简要概述

1.用测试化合物(100μL/ 96孔板或25μL/ 384孔板)制备细胞(样品)
2.加入等体积的ATP工作溶液(100μL/ 96孔板或25μL/ 384孔板)
3.在室温下孵育10-20分钟
4.监测发光强度

 

制备工作溶液

1.将10 mL反应缓冲液(组分C)转移到ATP传感器(组分B)中并充分混合。

2.将20μLATP监测酶(组分A)加入到组分B + C的瓶中并充分混合以制备ATP工作溶液。 注意:避免来自外源生物来源的潜在ATP污染。

有关细胞样品制备的指南(点击查看)

 

样品实验方案

1.运行ATP测定:

1.1用测试化合物处理细胞(或样品),在96孔板中加入10μL10X化合物,或在所需化合物缓冲液中加入5μL5X化合物用于384孔板。对于空白孔(没有细胞的培养基),加入相应量的化合物缓冲液。

1.2将细胞板在37℃,5%CO 2培养箱中孵育所需的一段时间,例如24,48或96小时。

1.3向每个孔中加入100μL(96孔板)或25μL(384孔板)的ATP工作溶液。

1.4在室温下孵育10-20分钟。

1.5用标准发光计监测发光强度。

 

2.生成标准ATP校准曲线:

        如果需要计算样品中ATP的绝对量,则应与上述分析一起生成ATP标准曲线。

2.1通过包含不含ATP的样品(作为对照)在含有0.1%BSA的PBS缓冲液中制备ATP系列稀释液以测量背景发光。注意:通常ATP浓度范围为0.1 nM至1μM是合适的。

2.2将相同量的稀释的ATP溶液加入空板中(对于96孔板为100μL或对于384孔板为25μL)。

2.3加入100μL/孔(96孔板)或25μL/孔(384孔板)的ATP工作溶液。

2.4将反应混合物在室温下孵育10至20分钟。

2.5用标准发光计记录发光强度。

2.6生成ATP标准曲线。

 

参考文献

High throughput cell-based assay for identification of glycolate oxidase inhibitors as a potential treatment for Primary Hyperoxaluria Type 1
Authors: Mengqiao Wang, Miao Xu, Yan Long, Sonia Fargue, Noel Southall, Xin Hu, John C McKew, Christopher J Danpure, Wei Zheng
Journal: Scientific Reports (2016)

NT1014, a novel biguanide, inhibits ovarian cancer growth in vitro and in vivo
Authors: Lu Zhang, Jianjun Han, Amanda L Jackson, Leslie N Clark, Joshua Kilgore, Hui Guo, Nick Livingston, Kenneth Batchelor, Yajie Yin, Timothy P Gilliam
Journal: Journal of Hematology & Oncology (2016): 91

The Different Effects of Atorvastatin and Pravastatin on Cell Death and PARP Activity in Pancreatic NIT-1 Cells
Authors: Ya-Hui Chen, Yi-Chun Chen, Chin-San Liu, Ming-Chia Hsieh
Journal: Journal of Diabetes Research (2016)

BPA-induced DNA hypermethylation of the master mitochondrial gene PGC-1α contributes to cardiomyopathy in male rats
Authors: Ying Jiang, Wei Xia, Jie Yang, Yingshuang Zhu, Huailong Chang, Juan Liu, Wenqian Huo, Bing Xu, Xi Chen, Yuanyuan Li
Journal: Toxicology (2015): 21–31

Glutamine promotes ovarian cancer cell proliferation through the mTOR/S6 pathway
Authors: Lingqin Yuan, Xiugui Sheng, Adam K Willson, Dario R Roque, Jessica E Stine, Hui Guo, Hannah M Jones, Chunxiao Zhou, Victoria L Bae-Jump
Journal: Endocrine-related cancer (2015): 577–591

JQ1 suppresses tumor growth through downregulating LDHA in ovarian cancer
Authors: Haifeng Qiu, Amanda L Jackson, Joshua E Kilgore, Yan Zhong, Leo Li-Ying Chan, Paola A Gehrig, Chunxiao Zhou, Victoria L Bae-Jump
Journal: Oncotarget (2015): 6915

Loss of histone deacetylase Hdac1 disrupts metabolic processes in intestinal epithelial cells
Authors: Alexis Gonneaud, Naomie Turgeon, Frančois-Michel Boisvert, Frančois Boudreau, Claude Asselin
Journal: FEBS letters (2015): 2776–2783

A neutrophil intrinsic impairment affecting Rab27a and degranulation in cystic fibrosis is corrected by CFTR potentiator therapy
Authors: Kerstin Pohl, Elaine Hayes, Joanne Keenan, Michael Henry, Paula Meleady, Kevin Molloy, Bakr Jundi, David A Bergin, Cormac McCarthy, Oliver J McElvaney
Journal: Blood (2014): 999–1009

Fluorescence lifetime imaging unravels C. trachomatis metabolism and its crosstalk with the host cell
Authors: Márta Szaszák, Philipp Steven, Kensuke Shima, Regina Orzekowsky-Schroder, Gereon Huttmann, Inke R Konig, Werner Solbach, Jan Rupp
Journal: PLoS pathogens (2011): e1002108

G protein coupled receptor kinase 2 interacting protein 1 (GIT1) is a novel regulator of mitochondrial biogenesis in heart
Authors: Jinjiang Pang, Xiangbin Xu, Michael R Getman, Xi Shi, Stephen L Belmonte, Heidi Michaloski, Amy Mohan, Burns C Blaxall, Bradford C Berk
Journal: Journal of molecular and cellular cardiology (2011): 769–776

 

相关产品

产品名称 货号
PhosphoWorks 发光法ATP检测试剂盒 *温和发光* Cat#21609
PhosphoWorks 发光法ATP检测试剂盒 *明亮发光* Cat#21610
PhosphoWorks 荧光ATP检测试剂盒 Cat#21620

说明书
PhosphoWorks 发光法ATP检测试剂盒* DTT – Free*.pdf

荧光标记的ATP-TAMRA conjugate [5-TAMRA-eda-ATP](停产) 货号13607-AAT Bioquest荧光染料

上海金畔生物科技有限公司代理AAT Bioquest荧光染料全线产品,欢迎访问AAT Bioquest荧光染料官网了解更多信息。

荧光标记的ATP-TAMRA conjugate [5-TAMRA-eda-ATP](停产)

荧光标记的ATP-TAMRA conjugate [5-TAMRA-eda-ATP](停产)

荧光标记的ATP-TAMRA conjugate [5-TAMRA-eda-ATP](停产)    货号13607 货号 13607 存储条件 在零下15度以下保存, 避免光照
规格 100 nmol 价格 0
Ex (nm) 552 Em (nm) 578
分子量 1128.13 溶剂 DMSO
产品详细介绍

简要概述

产品基本信息

货号:13607

产品名称:荧光标记的ATP-TAMRA conjugate [5-TAMRA-eda-ATP]

规格:100 nmol

储存条件:-15℃避光防潮

保质期:24个月

 

产品物理化学光谱特性

分子量:1128.13

溶剂:DMSO

激发波长(nm):544

发射波长(nm):575

 

产品介绍

荧光标记的ATP-TAMRA conjugate [5-TAMRA-eda-ATP]是美国AAT Bioquest生产的荧光染料,荧光标记的ATP示踪剂可用于监测ATP结合蛋白和其他生物分子。 ATP-TAMRA示踪剂被证明与ATPase结合,后者是一类酶,可催化ATP分解为ADP和游离磷酸根离子。金畔生物是AAT Bioquest的中国代理商,为您提供最优质的荧光标记的ATP-TAMRA conjugate [5-TAMRA-eda-ATP]。 

点击查看光谱

 

参考文献

Deciphering the catalysis-associated conformational changes of human adenylate kinase 1 with single-molecule spectroscopy
Authors: Lin CY, Huang JY, Lo LW.
Journal: J Phys Chem B (2013): 13947

The effect of NBD-Cl in nucleotide-binding of the major subunit alpha and B of the motor proteins F1FO ATP synthase and A1AO ATP synthase
Authors: Hunke C, Tadwal VS, Manimekalai MS, Roessle M, Gruber G.
Journal: J Bioenerg Biomembr (2010): 1

The Escherichia coli PriA helicase-double-stranded DNA complex: location of the strong DNA-binding subsite on the helicase domain of the protein and the affinity control by the two nucleotide-binding sites of the enzyme
Authors: Szymanski MR, Jezewska MJ, Bujalowski W.
Journal: J Mol Biol (2010): 344

ATP/ADP binding to a novel nucleotide binding domain of the reticulocyte-binding protein Py235 of Plasmodium yoelii
Authors: Ramalingam JK, Hunke C, Gao X, Gruber G, Preiser PR.
Journal: J Biol Chem (2008): 36386

Reversal of ADP-mediated aggregation of adenosine kinase by cyclophilin leads to its reactivation
Authors: Sen B, Chakraborty A, Datta R, Bhattacharyya D, Datta AK.
Journal: Biochemistry (2006): 263

ATPase mechanism of Eg5 in the absence of microtubules: insight into microtubule activation and allosteric inhibition by monastrol
Authors: Cochran JC, Gilbert SP.
Journal: Biochemistry (2005): 16633

Ca2+ binding to sarcoplasmic reticulum ATPase phosphorylated by Pi reveals four thapsigargin-sensitive Ca2+ sites in the presence of ADP
Authors: Vieyra A, Mintz E, Lowe J, Guillain F.
Journal: Biochim Biophys Acta (2004): 103

Evidence for proximal cysteine and lysine residues at or near the active site of arginine kinase of Stichopus japonicus
Authors: Guo Q, Chen B, Wang X.
Journal: Biochemistry (Mosc) (2004): 1336

Conformational dynamics of DnaB helicase upon DNA and nucleotide binding: analysis by intrinsic tryptophan fluorescence quenching
Authors: Flowers S, Biswas EE, Biswas SB.
Journal: Biochemistry (2003): 1910

D1 ring is stable and nucleotide-independent, whereas D2 ring undergoes major conformational changes during the ATPase cycle of p97-VCP
Authors: Wang Q, Song C, Yang X, Li CC.
Journal: J Biol Chem (2003): 32784

说明书
荧光标记的ATP-TAMRA conjugate [5-TAMRA-eda-ATP](停产).pdf

PhosphoWorks 发光测定ATP测定试剂盒* Bright Glow * 货号21621-AAT Bioquest荧光染料

上海金畔生物科技有限公司代理AAT Bioquest荧光染料全线产品,欢迎访问AAT Bioquest荧光染料官网了解更多信息。

PhosphoWorks 发光测定ATP测定试剂盒* Bright Glow *

PhosphoWorks 发光测定ATP测定试剂盒* Bright Glow *

PhosphoWorks 发光测定ATP测定试剂盒* Bright Glow *    货号21621 货号 21621 存储条件 在零下15度以下保存, 避免光照
规格 10 Plates 价格 12588
Ex (nm) Em (nm)
分子量 溶剂
产品详细介绍

简要概述

PhosphoWorks 荧光ATP检测试剂盒是美国AAT Bioquest生产的用于检测ATP的试剂盒,三磷酸腺苷(ATP)在细胞能量,代谢调节和细胞信号传导中起着基本作用。它被称为细胞内能量转移的“货币分子单位”,以驱动活细胞中的许多过程和化学合成。 ATP也是细胞通讯的信号分子,在DNA和RNA合成中起重要作用。 AAT Bioquest提供了多种生物发光测定试剂盒,可通过重组萤火虫荧光素酶(目录号21610和21609)确定ATP的纳摩尔(nM)范围。这些试剂盒需要酶标仪,通常用于细胞活力或细胞毒性测定。 PhosphoWorks 荧光ATP分析试剂盒基于一系列ATP诱导的酶偶联反应,产生过氧化氢,该过氧化氢用我们的Amplite Red底物通过分光光度法定量。该测定法可在100 µL反应体积中检测到约0.4 µM ATP,而不受ADP和AMP的干扰。它为测定生物样品中的ATP水平提供了一种强大,简单且方便的测定方法。 PhosphoWorks 荧光ATP分析是我们基于荧光素酶的ATP分析试剂盒的补充。金畔生物是AAT Bioquest的中国代理商,为您提供最优质的PhosphoWorks 荧光ATP检测试剂盒。

 

适用仪器


发光酶标仪  
推荐孔板: 白色孔板

产品说明书

样品实验方案

简要概述

1.准备ATP工作溶液(50 µL)
2.添加ATP标准液或测试样品(50 µL)
3.在室温下孵育10-30分钟
4.监测Ex / Em = 540/590 nm(截止= 570 nm)的荧光增加

 

溶液配制

1.制备储备溶液

所有未使用的储备溶液应分为一次性使用的等分试样,并在制备后储存在-20°C下。 避免重复冻融循环。
1.1Amplite 红色底物储备液(200X):
将30 µL DMSO(组分E)加入小瓶AmpliteTM Red Substrate(组分A)中,制成200X Amplite Red Substrate储备液。

1.2ATP标准溶液(10mM):
将0.5 mL的ddH2O加入到ATP标准溶液(组分D)的小瓶中,制成10 mM ATP标准溶液。

 

2.制备标准溶液

ATP标准

将10 µL的10 mM ATP标准溶液添加到990 µL 1X PBS缓冲液中,以生成100 µM ATP标准溶液(AS7)。 取100 µM ATP标准溶液(AS7)并进行1:3连续稀释,以使用1X PBS缓冲液获得系列稀释的ATP标准溶液(AS6-AS1)。

 

3.制备工作溶液

3.1将5毫升测定缓冲液(组分C)添加到酶混合瓶(组分B)中,并充分混合。

3.2在Enzyme Mix瓶中加入25 µL 200X AmpliteTM Red Substrate储备液,充分混合以制成APT工作溶液。

 

样品示例及操作

表1.黑色96孔微孔板中ATP标准品和测试样品的布局。 AS = ATP标准品(AS1-AS7,0.14至100 µM),BL =空白对照,TS =测试样品。

BL BL TS TS
AS1 AS1
AS2 AS2
AS3 AS3    
AS4 AS4    
AS5 AS5    
AS6 AS6    
AS7 AS7    

表2.每个孔的试剂组成

容积 试剂
AS1-AS7 50ul 连续稀释(0.14至100 µM)
BL 50ul 1 X PBS缓冲液
TS 50ul 测试样品

1.根据表1和2中提供的布局,准备ATP标准品(AS),空白对照(BL)和测试样品(TS)。对于384孔板,每孔使用25 µL试剂,而不是50 µL。

2.将50 µL ATP工作溶液添加到ATP标准液,空白对照和测试样品的每个孔中,以使ATP总测定体积为100 µL /孔。 对于384孔板,将25 µL ATP工作溶液添加到每个孔中,总体积为50 µL /孔。

3.避光保存,室温下孵育反应10-30分钟。

4.使用荧光酶标仪在Ex / Em = 540/590 nm(截止= 570 nm)处监视荧光增加。

 

参考文献

ATP-based cell viability assay is superior to trypan blue exclusion and XTT assay in measuring cytotoxicity of anticancer drugs Taxol and Imatinib, and proteasome inhibitor MG-132 on human hepatoma cell line HepG2
Authors: E. Nowak
Journal: Clin Hemorheol Microcirc (2018): 327-336

High-content image analysis (HCIA) assay has the highest correlation with direct counting cell suspension compared to the ATP, WST-8 and Alamar blue assays for measurement of cytotoxicity
Authors: H. Tahara
Journal: J Pharmacol Toxicol Methods (2017): 92-99

High throughput cell-based assay for identification of glycolate oxidase inhibitors as a potential treatment for Primary Hyperoxaluria Type 1
Authors: Mengqiao Wang, Miao Xu, Yan Long, Sonia Fargue, Noel Southall, Xin Hu, John C McKew, Christopher J Danpure, Wei Zheng
Journal: Scientific Reports (2016)

NT1014, a novel biguanide, inhibits ovarian cancer growth in vitro and in vivo
Authors: Lu Zhang, Jianjun Han, Amanda L Jackson, Leslie N Clark, Joshua Kilgore, Hui Guo, Nick Livingston, Kenneth Batchelor, Yajie Yin, Timothy P Gilliam
Journal: Journal of Hematology & Oncology (2016): 91

The Different Effects of Atorvastatin and Pravastatin on Cell Death and PARP Activity in Pancreatic NIT-1 Cells
Authors: Ya-Hui Chen, Yi-Chun Chen, Chin-San Liu, Ming-Chia Hsieh
Journal: Journal of Diabetes Research (2016)

BPA-induced DNA hypermethylation of the master mitochondrial gene PGC-1α contributes to cardiomyopathy in male rats
Authors: Ying Jiang, Wei Xia, Jie Yang, Yingshuang Zhu, Huailong Chang, Juan Liu, Wenqian Huo, Bing Xu, Xi Chen, Yuanyuan Li
Journal: Toxicology (2015): 21–31

Combination of immunoprecipitation (IP)-ATP_Glo kinase assay and melanogenesis for the assessment of potent and safe PAK1-blockers in cell culture
Authors: B. C. Nguyen
Journal: Drug Discov Ther (2015): 289-95

Glutamine promotes ovarian cancer cell proliferation through the mTOR/S6 pathway
Authors: Lingqin Yuan, Xiugui Sheng, Adam K Willson, Dario R Roque, Jessica E Stine, Hui Guo, Hannah M Jones, Chunxiao Zhou, Victoria L Bae-Jump
Journal: Endocrine-related cancer (2015): 577–591

JQ1 suppresses tumor growth through downregulating LDHA in ovarian cancer
Authors: Haifeng Qiu, Amanda L Jackson, Joshua E Kilgore, Yan Zhong, Leo Li-Ying Chan, Paola A Gehrig, Chunxiao Zhou, Victoria L Bae-Jump
Journal: Oncotarget (2015): 6915

Loss of histone deacetylase Hdac1 disrupts metabolic processes in intestinal epithelial cells
Authors: Alexis Gonneaud, Naomie Turgeon, Frančois-Michel Boisvert, Frančois Boudreau, Claude Asselin
Journal: FEBS letters (2015): 2776–2783

 

参考文献

产品名称 货号
PhosphoWorks 比色ATP分析试剂盒 Cat#21617
PhosphoWorks 发光法ATP检测试剂盒 *温和发光* Cat#21609
PhosphoWorks 发光法ATP检测试剂盒 *明亮发光* Cat#21610

说明书
PhosphoWorks 发光测定ATP测定试剂盒* Bright Glow *.pdf