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当前位置: 首页 > 产品中心 > Other_kits > AAT Bioquest/Screen Quest™ Calbryte-590 Probenecid-Free and Wash-Free Calcium Assay Kit/36201/10 Plates
商品详细AAT Bioquest/Screen Quest™ Calbryte-590 Probenecid-Free and Wash-Free Calcium Assay Kit/36201/10 Plates
AAT Bioquest/Screen Quest™ Calbryte-590 Probenecid-Free and Wash-Free Calcium Assay Kit/36201/10 Plates
AAT Bioquest/Screen Quest™ Calbryte-590 Probenecid-Free and Wash-Free Calcium Assay Kit/36201/10 Plates
商品编号: 36201
品牌: aatbio
市场价: ¥19000.00
美元价: 11400.00
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
PrinterFriendlyVersion

Ex/Em(nm)573/588
MWN/A
CAS#N/A
SolventN/A
StorageF/D/L
CategoryGPCR
CalciumGPCRAssays
RelatedCalciumChannels
pHandIonIndicators
CalciumfluxassaysarethepreferredmethodsforscreeningGproteincoupledreceptors(GPCRs)indrugdiscovery.ScreenQuest™Calbryte-590™Probenecid-FreeandWash-FreeCalciumAssayKitprovidesthemostrobusthomogeneousredfluorescence-basedassayfordetectingtheintracellularcalciummobilization.CellsexpressingaGPCRofinterestthatsignalsthroughcalciumarepre-loadedwithourproprietaryCalbryte-590AMwhichcancrosscellmembrane.Calbryte-590AMisthebrightestcalciumindicatoravailableforHTSscreening.Onceinsidethecell,thelipophilicblockinggroupsofCalbryte-590AMarecleavedbynon-specificcellesterase,resultinginanegativelychargedfluorescentdyethatstaysinsidecells,anditsfluorescenceisgreatlyenhanceduponbindingtocalcium.Whencellsstimulatedwithscreeningcompounds,thereceptorsignalsreleaseofintracellularcalcium,whichgreatlyincreasethefluorescenceofCalbryte-590AM.Thecharacteristicsofitsexcellentcellretention,highsensitivity,and100-250timesfluorescenceincreases(whenitformscomplexeswithcalcium)makeCalbryte-590AManidealindicatorformeasurementofcellularcalcium.Calbryte-590AMistheonlyredcalciumdyethatdoesnotrequireprobenecidtoimprovecellularretention.ThisScreenQuest™Calbryte-590™Probenecid-FreeandWash-FreeCalciumAssayKitprovidesthemostoptimizedassaymethodformonitoringGPCRsandcalciumchannelswithfragileordifficultcelllines.Comparedtothegreenfluorescence-basedcalciumassays,thisassaykithaslessinterferencefromthecoloredcompoundsfromacompoundlibrary.ItisalsocompatIBLewithGFPcelllinesforhighcontentanalysis.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. PrepareCalbryte™590AMdye-loADIngsolution:
    1. Thawallthekitcomponentsatroomtemperaturebeforeuse.
    2. MakeCalbryte™590AMstocksolution:Add20µL(forCat#36200)or200µL(forCat#36201and36202)ofDMSOintothevialofCalbryte™590AM(ComponentA),andmixthemwell.
      Note:20µLofCalbryte™590AMstocksolutionisenoughforoneplate.UnusedCalbryte™590AMstocksolutioncanbealiquotedandstoredat<-20=""oc=""for=""more=""than=""one=""month=""if=""the=""tubes=""are=""sealed=""tightly.=""protect=""from=""light=""and=""avoid=""repeated=""freeze-thaw="">
    3. Make1Xassaybuffer:Mix9mLofHHBS(ComponentC,notincludedinthekitCat#36202)with1mLof10XPluronicF127Plus(ComponentB),andmixthemwell.
    4. MakeCalbryte™590AMdye-loadingsolutionforonecellplate:Add20µLofCalbryte™590AMstocksolution(fromStep2.2)into10mLof1Xassaybuffer(fromStep2.3),andmixthemwell.Thisworkingsolutionisstableforatleast2hoursatroomtemperature.

  3. Runcalciumassay:
    1. Add100µL/well(96-wellplate)or25µL/well(384-wellplate)ofCalbryte™590AMdye-loadingsolution(fromStep2.4)intothecellplate.
    2. Incubatethedye-loadingplateinacellincubatorfor~60minutes,andthenincubatetheplateatroomtemperatureforanother15minutes.
      Note1:Iftheassayrequires37oC,performtheexperimentimmediatelywithoutfurtherroomtemperatureincubation.
      Note2:Ifthecellscanfunctionwellatroomtemperatureforlongertime,incubatethecellplateatroomtemperaturefor1hour.
    3. PreparethecompoundplatewithHHBSoryourdesiredbuffer.
    4. RunthecalciumfluxassaybymonitoringthefluorescenceintensitywithTRITCFilter(orEx/Em=540/590nm).
References&Citations
CitationExplorer

CalreticulinregulatesTGF-β1-inducedepithelialmesenchymaltransitionthroughmodulatingSmadsignalingandcalciumsignaling
Authors:YanjiaoWu,XiaoliXu,LunkunMa,QianYi,WeichaoSun,LilingTang
Journal:TheInternationalJournalofBiochemistry&CellBIOLOGy(2017)

Dexmedetomidinereduceshypoxia/reoxygenationinjurybyregulatingmitochondrialfissioninrathippocampalneurons
Authors:JiaLiu,QingDu,HeZhu,YuLi,MaodongLiu,ShoushuiYu,ShileiWang
Journal:IntJClinExpMed(2017):6861--6868

Monosialoganglioside1mayalleviateneurotoxicityinducedbypropofolcombinedwithremifentanilinneuralstemcells
Authors:JiangLu,Xue-qinYao,XinLuo,YuWang,SookjaKimChung,He-xinTang,ChiWaiCheung,Xian-yuWang,ChenMeng,QingLi
Journal:NeuralRegenerationResearch(2017):945

Obtainingspontaneouslybeatingcardiomyocyte-likecellsfromadipose-derivedstromalvascularfractionsculturedonenzyme-crosslinkedgelatinhydrogels
Authors:GangYang,ZhenghuaXiao,XiaomeiRen,HaiyanLong,KunlongMa,HongQian,YingqiangGuo
Journal:ScientificReports(2017):41781

Di(2-ethylhexyl)phthalate-inducedapoptosisinratINS-1cellsisdependentonactivationofendoplasmicreticulumstressandsuppressionofantioxidantprotection
Authors:XiaSun,YiLin,QianshengHuang,JunpengShi,LingQiu,MeiKang,YajieChen,ChaoFang,TingYe,SijunDong
Journal:Journalofcellularandmolecularmedicine(2015):581--594

Theeffectofmitochondrialcalciumuniporteronmitochondrialfissioninhippocampuscellsischemia/reperfusioninjury
Authors:LantaoZhao,ShuhongLi,ShileiWang,NingYu,JiaLiu
Journal:Biochemicalandbiophysicalresearchcommunications(2015):537--542

FungusinducesthereleaseofIL-8inhumancornealepithelialcells,viaDectin-1-mediatedproteinkinaseCpathways.
Authors:Xu-DongPeng,Gui-QiuZhao,JingLin,NanJiang,QiangXu,Cheng-ChengZhu,Jain-QiuQu,LinCong,HuiLi
Journal:Internationaljournalofophthalmology(2014):441--447

Propofolandremifentanilatmoderateandhighconcentrationsaffectproliferationanddifferentiationofneuralstem/Progenitorcells
Authors:QingLi,JiangLu,XianyuWang
Journal:Neuralregenerationresearch(2014):2002

Roleofmitochondrialcalciumuniporterinregulatingmitochondrialfissioninthecerebralcortexesoflivingrats
Authors:NanLiang,PengWang,ShileiWang,ShuhongLi,YuLi,JinyingWang,MinWang
Journal:JournalofNeuralTransmission(2014):593--600

Increasedexpressionofcelladhesionmolecule1bymastcellsasacauseofenhancednerve--mastcellinteractioninahapten-inducedmousemodelofatopicdermatitis
Authors:MHagiyama,TInoue,TFuruno,TIino,SItami,MNakanishi,HAsada,YHosokawa,AIto
Journal:BritishJournalofDermatology(2013):771--778


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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