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当前位置: 首页 > 产品中心 > Other_kits > AAT Bioquest/Screen Quest™ Fluo-4 No Wash Calcium Assay Kit/36326/100 Plates
商品详细AAT Bioquest/Screen Quest™ Fluo-4 No Wash Calcium Assay Kit/36326/100 Plates
AAT Bioquest/Screen Quest™ Fluo-4 No Wash Calcium Assay Kit/36326/100 Plates
AAT Bioquest/Screen Quest™ Fluo-4 No Wash Calcium Assay Kit/36326/100 Plates
商品编号: 36326
品牌: aatbio
市场价: ¥59000.00
美元价: 35400.00
产地: 美国(厂家直采)
公司:
产品分类: 其它检测试剂盒
公司分类: Other_kits
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)490/525
MWN/A
CAS#N/A
SolventN/A
StorageF/D/L
CategoryGPCR
CalciumGPCRAssays
RelatedCalciumChannels
pHandIonIndicators
CalciumfluxassaysarepreferredmethodsindrugdiscoveryforscreeningGproteincoupledreceptors(GPCR).ScreenQuest™Fluo-4NoWashCalciumAssayKitprovidesahomogeneousfluorescence-basedassayfordetectingtheintracellularcalciummobilization.CellsexpressingaGPCRofinterestthatsignalsthroughcalciumarepre-loadedwithFluo-4AMwhichcancrosscellmembrane.Onceinsidethecell,thelipophilicblockinggroupsofFluo-4AMarecleavedbynon-specificcellesterase,resultinginanegativelychargedFluo-4dyethatstaysinsidecells,anditsfluorescenceisgreatlyenhanceduponbindingtocalcium.Whencellsstimulatedwithscreeningcompounds,thereceptorsignalsreleaseofintracellularcalcium,whichgreatlyincreasethefluorescenceofFluo-4.ThisScreenQuestFluo-4NoWashCalciumAssayKitprovidesanoptimizedassaymethodformonitoringG-protein-coupledreceptors(GPCRs)andcalciumchannels.Theassaycanbeperformedinaconvenient96-wellor384-wellmicrotiter-plateformatandeasilyadaptedtoautomation.
SpectrumAdvancedSpectrumViewer

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Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds.
  1. Preparecells:
    1. Foradherentcells:Platecellsovernightingrowthmediumat40,000to80,000cells/well/100µLfora96-wellplateor10,000to20,000cells/well/25µLfora384-wellplate.
    2. Fornon-adherentcells:CentrifugethecellsfromtheculturemediumandthensUSPendthecellpelletincellgrowthmediumorHHBSat125,000to250,000cells/well/100µLfora96-wellpoly-Dlysineplateor30,000to60,000cells/well/25µLfora384-wellpoly-Dlysineplate.Centrifugetheplateat800rpmfor2minuteswithbrakeoffpriortotheexperiments.
      Note:Eachcelllineshouldbeevaluatedontheindividualbasistodeterminetheoptimalcelldensityfortheintracellularcalciummobilization.

  2. PrepareFluo-4AMdye-loADIngsolution:
    1. Thawallthekitcomponentsatroomtemperaturebeforeuse.
    2. MakeFluo-4AMstocksolution:Add200µLofDMSOintothevialofFluo-4AM(ComponentA),andmixthemwell.
      Note:20µLofFluo-4NWstocksolutionisenoughforoneplate.UnusedFluo-4AMstocksolutioncanbealiquotedandstoredat<-20=""°c=""for=""more=""than=""one=""month=""if=""the=""tubes=""are=""sealed=""tightly.=""protect=""from=""light=""and=""avoid=""repeated=""freeze-thaw=""cycles.="">
    3. Make1Xassaybuffer:
      Make1Xassaybufferbyadding1mLof10XPluronic®F127Plus,(10mL,ComponentB)into9mLofHHBSbuffer(ComponentC),andmixthemwell.
      Note:10mLof1Xassaybufferisenoughforoneplate.Aliquotandstoreun-used10XPluronic®F127Plus(ComponentB)at<-20=""°c.=""protect=""from=""light=""and=""avoid=""repeated=""freeze-thaw="">
    4. MakeFluo-4AMdye-loadingsolutionforonecellplate:Add20µLofFluo-4NWstocksolution(fromStep2.2)into10mLof1Xassaybuffer(fromStep2.3),andmixthemwell.Thisworkingsolutionisstableforatleast2hoursatroomtemperature.

  3. Runcalciumassay:
    1. Add100µL/well(96-wellplate)or25µL/well(384-wellplate)ofFluo-4AMdye-loadingsolution(fromStep2.4)intothecellplate.
    2. Incubatethedye-loadingplateinacellincubatorfor1hour,andthenincubatetheplateatroomtemperatureforanother15to30minutes.
      Note1:Iftheassayrequires37°C,performtheexperimentimmediatelywithoutfurtherroomtemperatureincubation.
    3. PreparethecompoundplatewithHHBSoryourdesiredbuffer.
    4. RunthecalciumfluxassaybymonitoringthefluorescenceintensityatEx/Em=490/525nm.
References&Citations
CitationExplorer

LysophosphatidylcholineanditsphosphorothioateanaloguespotentiateinsulinsecretionviaGPR40(FFAR1),GPR55andGPR119receptorsinadifferentmanner
Authors:AnnaDrzazga,HjaltiKristinsson,MaciejSalaga,HubertZatorski,MariaKoziolkiewicz,EdytaGendaszewska-Darmach,PeterBergsten
Journal:MolecularandCellularEndocrinology(2017)


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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