SunRed 醋酸 细胞活性检测-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。
SunRed 醋酸 细胞活性检测 价格 1386
产品规格

25 mg

产品货号

SunRed 醋酸 细胞活性检测

产品参数
Ex (nm) 653 Em (nm) 661
分子量 ~350 溶剂 DMSO
存储条件 在零下15度以下保存, 避免光照
产品概述

SunRed 醋酸 细胞活性检测是美国AAT Bioquest生产的用于细胞活性检测的试剂盒,酯酶可催化水解SunRed 乙酸盐(SRA)产生Sun Red荧光基团,该荧光基团在633nm处可以被激发,发射光波长在~660nm处。Sun Red荧光基团可以用Cy5滤光片检测,大多数商业荧光仪器中都装有Cy5滤光片。尽管Sun Red在633nm处被激发,~660nm发出红光;但是SRA在633nm处的光吸收很小,在没有红光发射的条件下,这使SRA成为灵敏的NIR(近红外)酯酶底物之一。在流行的荧光色彩应用于其它细胞功能分析时,SRA提供另一种颜色用于可行性细胞的分析。SRA是一个非荧光性的疏水性化合物,它可以通过细胞膜,在细胞内酯酶水解乙酸盐基团作用下,产生高强度荧光的产物Sun Red。Sun Red分子在完整细胞膜结构的细胞内积累至产生足够深的红色荧光,因此可以用作细胞的标签。细胞膜结构不完整或者代谢活性弱的细胞,不能够积累荧光产物,因此不显现深色的红色荧光。SRA可以与绿色荧光染料试剂(例如FITC或者Alexa Fluor 488标记的抗体)结合使用。金畔生物是AAT Bioquest的中国代理商,为您提供优质的SunRed 醋酸 细胞活性检测。

点击查看光谱

 

适用仪器


荧光显微镜  
Ex: 620nm
Em: 660nm
推荐孔板: 黑色透明底板
滤波片: Cy5滤波片组
   
荧光酶标仪  
Ex: 620nm
Em: 660nm
Cutoff: 640nm
推荐孔板: 纯黑色孔板
实验方案

样品实验方案

简要概述

1.用测试化合物制备细胞
2.去除培养基
3.添加SunRed 醋酸酯工作溶液(100μL/孔/ 96孔板或25μL/孔/ 384孔板)
4.在37°C,5%CO2培养箱中孵育1小时
5.用HHBS清洗并更换工作溶液
6.读取Ex / Em = 620/660 nm处的荧光强度(截止640 nm)

 

溶液制备 

1.储备溶液

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。
1.1 SunRed 醋酸盐原液(2至10 mM)
准备2至10 mM的SunRed 醋酸盐在DMSO中的储备溶液。 应立即使用原液。

  

2.工作溶液

SunRed 醋酸盐工作溶液:
在实验前,在含有20 mM Hepes缓冲液(HHBS)或您选择的缓冲液的1X Hank盐溶液中,以5至10μM制备SunRed 醋酸盐工作溶液。

点击查看样品制备方案

 

样品分析

1.从细胞中取出培养基。

2.加入100μL/孔(96孔板)或25μL/孔(384孔板)的SunRed 乙酸盐工作溶液。

3.将SunRed 醋酸盐工作溶液板在室温或37°C下孵育1小时,避光。 适当的孵育时间取决于所用的单个细胞类型和细胞浓度。 优化每个实验的孵育时间。 对于非粘附细胞,建议在孵育后以800rpm离心细胞板2分钟。

4.监测Ex / Em = 620 / 660nm处的荧光强度(截止640nm)。

 

参考文献

Neuropathy target esterase is required for adult vertebrate axon maintenance
Authors: Read DJ, Li Y, Chao MV, Cavanagh JB, Glynn P.
Journal: J Neurosci (2009): 11594

The secreted esterase of group a streptococcus is important for invasive skin infection and dissemination in mice
Authors: Zhu H, Liu M, Sumby P, Lei B.
Journal: Infect Immun (2009): 5225

Application of glutaraldehyde for the staining of esterase-active cells with carboxyfluorescein diacetate
Authors: Morono Y, Takano S, Miyanaga K, Tanji Y, Unno H, Hori K.
Journal: Biotechnol Lett (2004): 379

Evaluation of fitness components in strains of Drosophila mulleri carrying different genotypes for an esterase
Authors: Lourenco MF, Ceron CR, Carareto CM.
Journal: Cytobios (2001): 125

Comparison of two in vitro activation systems for protoxicant organophosphorous esterase inhibitors
Authors: Barber D, Correll L, Ehrich M.
Journal: Toxicol Sci (1999): 16

Determination of growth and lysis kinetics in plant cell suspension cultures from the measurement of esterase release
Authors: Steward N, Martin R, Engasser JM, Goergen JL.
Journal: Biotechnol Bioeng (1999): 114

Purification and properties of an esterase from the yeast Saccharomyces cerevisiae and identification of the encoding gene
Authors: Degrassi G, Uotila L, Klima R, Venturi V.
Journal: Appl Environ Microbiol (1999): 3470

Organophosphorus neuropathy target esterase inhibitors selectively block outgrowth of neurite-like and cell processes in cultured cells
Authors: Li W, Casida JE.
Journal: Toxicol Lett (1998): 139

Causes and consequences of esterase 6 enzyme activity variation in pre-adult Drosophila melanogaster
Authors: Oakeshott JG, Saad M, Game AY, Healy MJ.
Journal: Heredity (1994): 160

[Esterase-6 gene-enzyme system and resistance of Drosophila to increased temperature]
Authors: Totskii VN, Eserkepova EV, Dzhan ZU.
Journal: Genetika (1994): 342

D-荧光素醋酸-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。
D-荧光素醋酸价格 2823
产品规格

5 mg

产品货号

D-荧光素醋酸

产品参数
Ex (nm) 385 Em (nm) 529
分子量 322.36 溶剂 DMSO
存储条件 在零下15度以下保存, 避免光照
产品概述

产品基本信息

产品名称:D-荧光素醋酸

储存条件:-15℃避光防潮

保质期:12个月

 

产品物理化学光谱特性

分子量:322.36

溶剂:DMSO

激发波长(nm):328

发射波长(nm):533

 

产品介绍

D-荧光素醋酸是美国AAT Bioquest生产的荧光染料,D-荧光素乙酯含有在羧基位置连接的甲基部分,因此防止它被萤火虫荧光素酶识别。 然而,酯通过脂肪酶活性有效地水解,并且所得的D-荧光素被荧光素酶很好地识别。 D-荧光素甲酯是用于在均相测定中或在与荧光素酶及其辅因子组合的细胞裂解物样品中化学发光测量脂肪酶活性的敏感底物。金畔生物是AAT Bioquest的中国代理商,为您提供优质的D-荧光素醋酸。 

Luc的Ex/Em及其他相关内容链接:Spectral properties of luciferin and EGFP. A: Normalized excitation and… | Download Scientific Diagram (researchgate.net)

金畔问号:荧光素酶到底该如何检测?

 

试剂应用文献

C3-Luc Cells Are an Excellent Model for Evaluation of Cellular Immunity following HPV16L1 Vaccination
Authors: Li-Li Li, He-Rong Wang, Zhi-Yi Zhou, Jing Luo, Xiao-Li Wang, Xiang-Qian Xiao, Yu-Bai Zhou, Yi Zeng
Journal: PloS one (2016): e0149748

Genome-wide microRNA analysis identifies miR-188-3p as novel prognostic marker and molecular factor involved in colorectal carcinogenesis
Authors: Martin Pichler, Verena Stiegelbauer, Petra Vychytilova-Faltejskova, Cristina Ivan, Hui Ling, Elke Winter, Xinna Zhang, Matthew Goblirsch, Annika Wulf-Goldenberg, Masahisa Ohtsuka
Journal: American Association for Cancer Research (2016): clincanres–0497

Identification of a Novel Protein Kinase A Inhibitor by Bioluminescence-Based Screening
Authors: Tetsuya Ishimoto, Kenji Azechi, Hisashi Mori
Journal: Biological and Pharmaceutical Bulletin (2015): 1969–1974

Discovery of novel adenylyl cyclase inhibitor by cell-based screening
Authors: Hiroki Mano, Tetsuya Ishimoto, Takuya Okada, Naoki Toyooka, Hisashi Mori
Journal: Biological and Pharmaceutical Bulletin (2014): 1689–1693

 

参考文献

Mutagenesis of solvent-exposed amino acids in Photinus pyralis luciferase improves stability and pH-tolerance
Authors: Law GH, G and elman OA, Tisi LC, Lowe CR, Murray JA.
Journal: Biochem J (2006): 305
 
Determination of p-glycoprotein ATPase activity using luciferase
Authors: Matsunaga T, Kose E, Yasuda S, Ise H, Ikeda U, Ohmori S.
Journal: Biol Pharm Bull (2006): 560
 
Expression of firefly luciferase in Candida albicans and its use in the selection of stable transformants
Authors: Doyle TC, Nawotka KA, Purchio AF, Akin AR, Francis KP, Contag PR.
Journal: Microb Pathog (2006): 69
 
PET imaging and optical imaging with D-luciferin [11C]methyl ester and D-luciferin [11C]methyl ether of luciferase gene expression in tumor xenografts of living mice
Authors: Wang JQ, Pollok KE, Cai S, Stantz KM, Hutchins GD, Zheng QH.
Journal: Bioorg Med Chem Lett (2006): 331
 
Bioluminescence of monolayers of firefly luciferase immobilized on graphite
Authors: Palomba S, Berovic N, Palmer RE.
Journal: Langmuir (2006): 5451
 
Bovine serum albumin displays luciferase-like activity in presence of luciferyl adenylate: insights on the origin of protoluciferase activity and bioluminescence colours
Authors: Viviani VR, Ohmiya Y.
Journal: Luminescence (2006): 262
 
Noninvasive bioluminescence imaging of luciferase expressing intracranial U87 xenografts: correlation with magnetic resonance imaging determined tumor volume and longitudinal use in assessing tumor growth and antiangiogenic treatment effect
Authors: Szentirmai O, Baker CH, Lin N, Szucs S, Takahashi M, Kiryu S, Kung AL, Mulligan RC, Carter BS.
Journal: Neurosurgery (2006): 365
 
Two different domains of the luciferase gene in the heterotrophic dinoflagellate Noctiluca scintillans occur as two separate genes in photosynthetic species
Authors: Liu L, Hastings JW.
Journal: Proc Natl Acad Sci U S A. (2006)
 
Effect of different salts and detergents on luciferin-luciferase luminescence of the enchytraeid Fridericia heliota
Authors: Rodionova NS, Petushkov VN.
Journal: J Photochem Photobiol B (2006): 123
 
Evaluation of firefly luciferase bioluminescence mediated photodynamic toxicity in cancer cells
Authors: Schipper ML, Patel MR, Gambhir SS.
Journal: Mol Imaging Biol (2006): 218