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当前位置: 首页 > 产品中心 > Other_biological_dyes > AAT Bioquest/DiSBAC2(5) [Bis-(1,3-diethylthiobarbituric acid)pentamethine oxonol]/21415/25 mg
商品详细AAT Bioquest/DiSBAC2(5) [Bis-(1,3-diethylthiobarbituric acid)pentamethine oxonol]/21415/25 mg
AAT Bioquest/DiSBAC2(5) [Bis-(1,3-diethylthiobarbituric acid)pentamethine oxonol]/21415/25 mg
AAT Bioquest/DiSBAC2(5) [Bis-(1,3-diethylthiobarbituric acid)pentamethine oxonol]/21415/25 mg
商品编号: 21415
品牌: aatbio
市场价: ¥42060.00
美元价: 25236.00
产地: 美国(厂家直采)
公司:
产品分类: 其他生物染料
公司分类: Other_biological_dyes
联系Q Q: 3392242852
电话号码: 4000-520-616
电子邮箱: info@ebiomall.com
商品介绍
Overview
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Ex/Em(nm)636/660
MW462.59
CAS#N/A
SolventDMSO
StorageF/D/L
CategoryIonChannels
MembranePotentials
RelatedNeurologicalStains
BiochemicalAssays
DiSBAC2(5)isasensitiveslow-responsemembranepotentialprobethatiswidelyusedformeasuringmembranepotentialsofmanyBIOLOGicalsystems.Ingeneral,slow-responseprobesexhibitpotential-dependentchangesintheirtransmembranedistributionthatareaccompaniedbyafluorescencechange.Themagnitudeoftheiropticalresponsesismuchlargerthanthatoffast-responseprobes(typicallya1%fluorescencechangepermV).Slow-responseprobes,whichincludecationiccarbocyanines,rhodaminesandanionicoxonols,aresuitablefordetectingchangesinaveragemembranepotentialsofnonexcitablecellscausedbyrespiratoryactivity,ion-channelpermeABIlity,drugbindingandotherfactors.

Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds.

1.Preparecells:

1.1   Foradherentcells:Platecellsovernightingrowthmediumat40,000to80,000cells/well/100µLfor96-wellplatesor10,000to20,000cells/well/25µLfor384-wellplates.

 

1.2   Fornon-adherentcells:CentrifugethecellsfromtheculturemediumandthensUSPendthecellpelletsinequalamountofHHBSandMPdye-loADIngsolution(seeStep2.2below)at125,000to250,000cells/well/100µLfor96-wellpoly-Dlysineplatesor30,000to60,000cells/well/25µLfor384-wellpoly-Dlysineplates.Centrifugetheplatesat800rpmfor2minuteswithbrakeoffbeforetheexperiments.

Note:Eachcelllineshouldbeevaluatedonanindividualbasistodeterminetheoptimalcelldensityfortheintracellularcalciummobilization.

 

2.PrepareDiSBAC2(3)dye-loadingsolution(for1plate):

2.1   Preparea10to30mMstocksolutionofDiSBAC2(3)inhigh-quality,anhydrousDMSO.Thestocksolutionshouldbeusedpromptly;anyremainingsolutionneedbealiquotedandfrozenat<-20oC.

Note:Avoidrepeatedfreeze-thawcycles,andprotectfromlight.

2.2   Preparea2XDiSBAC2(3)dye-loadingsolution:Onthedayoftheexperiment,eitherdissolveDiSBAC2(3)solidinDMSOorthawanaliquotoftheDiSBAC2(3)stocksolutiontoroomtemperature.Preparea2Xworkingsolutionof20to40µMinHanksand20mMHepesbuffer(HHBS)orbufferofyourchoice,pH7with0.04%to0.08%Pluronic®F-127(Cat.#20053)and2mMTrypanRedPlus™(Cat.#2456).Mixthemwellbyvotexing.Thisworkingsolutionisstableforatleast2hoursatroomtemperature.

 

3.RunMembranePotentialAssay:

3.1   Add100µL/well(96-wellplate)or25µL/well(384-wellplate)DiSBAC2(3)dye-loadingsolution(fromStep2.2)intothecellplate.

Note1:Ifyourscreencompoundsinterferewithgrowthmediumandserumfactors,replacethegrowthmediumwithequalvolumeofHHBSbufferbeforeaddingtheDiSBAC2(3)dye-loadingsolution.Alternatively,cellscanbegrowninserum-freeconditions.

Note2:DoNOTwashthecellsafterdyeloading.

 

3.2   Incubatethedye-loadingplateinacellincubatorfor30to60minutes.

Note:Insomecases,incubationatroomtemperaturefor30to60minmayworkbetter.

 

3.3   PreparethecompoundplatesbyusingHHBSoryourdesiredbuffer.

 

3.4   RunthemembranepotentialassaybymonitoringthefluorescenceintensityatEx/Em=540/590nm(Cat.# 21414).

Note:Itisimportanttorunthesignaltestbeforeyourexperiment.Differentinstrumentshavetheirownintensityrange.Adjustthesignaltestintensitytothelevelof10%to15%ofthemaximuminstrumentintensitycounts.Forexample,themaximumfluorescenceintensitycountforFLIPR-384is65,000,sotheinstrumentsettingsshouldbeadjustedtohaveitssignaltestintensityaround7,000to10,000.

References&Citations
CitationExplorer

SuppressionofKV7/KCNQpotassiumchannelenhancesneuronaldifferentiationofPC12cells
Authors:NajingZhou,ShaHuang,LiLi,DongyangHuang,YunliYan,XiaonaDu,HailinZhang
Journal:Neuroscience(2016):356--367


品牌介绍
美国AAT Bioquest Inc.(前身是ABD Bioquest,Inc.)是一家为从事生命科学研究、诊断研发及药物开发的科学家研发、生产和销售生物分析研究试剂和试剂盒的公司。公司致力于光谱学检测领域,包括吸收(颜色)、荧光和发光技术。AAT Bioquest的产品帮助全世界的科学家和生物医药研究者更好的了解生物化学、免疫学、细胞生物学和分子生物学等领域。AAT Bioquest会不断研发新产品,快速地丰富各个领域的产品。 1)提供反应探针和发光探针,生物素和端粒酶能够应用于标记药物小分子和生物聚合物,如蛋白、核酸以及其他碳水化合物; 2)研究并生产荧光和发光探针用于检测蛋白,核酸和活细胞; 3)不断推出新型的荧光和发光探针用于检测多种酶,特别是检测水解酶和氧化还原酶类; 4)致力于开发用于信号转导研究的试剂; 5)提供生理和神经探针,特别是钙离子指示剂和膜电位探针。
品牌分类
淋巴细胞信号传导 GPCR抗体 羰基活性生物素 细胞代谢 胺反应生物素 链霉亲和素结合物 生物素化抗IgGs 乙酰基特异性抗体 细胞骨架抗体
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