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主营:研究并生产荧光和发光探针,信号转导研究的试剂
℡ 4000-520-616
℡ 4000-520-616
AAT Bioquest/Calbryte™ 520 AM/20651/10x50 ug
产品编号:20651
市  场 价:¥5900.00
场      地:美国(厂家直采)
联系QQ:1570468124
电话号码:4000-520-616
邮      箱: info@ebiomall.com
美  元  价:$295.00
品      牌: AAT Bioquest
公      司:AAT Bioquest
公司分类:
AAT Bioquest/Calbryte™ 520 AM/20651/10x50 ug
商品介绍
Overview
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Ex/Em(nm)492/514
MW1090.90
CAS#N/A
SolventDMSO
StorageF/D/L
CategoryCellBIOLOGy
pHandIonIndicators
Related
TheintracellularcalciumfluxassayisawidelyusedmethodinmonitoringsignaltransductionpathwaysandhighthroughputscreeningofGprotein–coupledreceptors(GPCRs)andcalciumchanneltargets.FollowedbyFluo-3beingintroducedin1989,Fluo-4,Fluo-8andCal-520werelaterdevelopedwithimprovedsignal/backgroundratio,andbecamethewidelyusedCa2+indicatorsforconfocalmicroscopy,flowcytometryandhighthroughputscreeningapplications.However,therearestillafewsevereproblemswithFluo-4.Forexample,asforFluo-3,inallmostalltheintracellularcalciumassayswithFluo-4AM,probenecidisrequiredtopreventthecell-loadedFluo-4fromleakingoutofcells.TheuseofprobenecidwithFluo-4-basedcalciumassayscompromisestheassayresultssinceprobenecidiswell-documentedtohaveavarietyofcomplicatedcellulareffects.Calbryte™520,AMisanewfluorescentandcell-permeablecalciumindicator.LikeotherdyeAMcellloADIng,Calbryte™520AMesterisnon-fluorescentandoncegetsinsidecells,itishydrolyzedbyintracellularesteraseandgetsactivated.Theactivatedindicatorisapolarmoleculethatisnolongercapableoffreelydiffusingthroughcellmembrane,essentiallytrappedinsidecells.UponbindingCa2+ions,Calbryte™520producesbrightfluorescencesignalwithextremelyhighsignal/backgroundratio.Inaddition,Calbryte™520demonstratesgreatlyimprovedintracellularretention.IthastheidenticalexcitationandemissionwavelengthasFluo-4,thusthesameFluo-4assaysettingscanbereadilyappliedtoCalbryte™520-basedcalciumassays.Calbryte™520isanewgenerationoffluorescentindicatorsforthemeasurementofintracellularcalcium.Itsgreatlyimprovedsignal/backgroundratioandintracellularretentionpropertiesmakeCalbryte™520AMthemostrobustindicatorforevaluatingGPCRandcalciumchanneltargetsaswellasforscreeningtheiragoNISTsandantagonistsinlivecells.
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Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds.
  1. LoadCellswithCalbryte™520,Calbryte™590orCalbryte™630AMEsters:
    Calbryte™AMestersshouldbereconstitutedjustbeforeuseinanhydrousDMSO.TheDMSOstocksolutionsmaybestored(desiccated)at–20°Candprotectedfromlight.Undertheseconditions,AMestersshouldbestableforthreemonths.FollowingisourrecommendedprotocolforloadingCalbryte™520AM,Calbryte™590AMorCalbryte™630AMestersintolivecells.Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds.

    1. Preparea2to5mMstocksolutionofCalbryte™520AM,Calbryte™590AMorCalbryte™630AMestersinanhydrousDMSO.
    2. DissolveCalbryte™520AM,Calbryte™590AMorCalbryte™630AMinDMSOorthawanaliquotoftheindicatorstocksolutiontoroomtemperature.Prepareadyeworkingsolutionof10to20µMinHanksandHepesbuffer(HHBS)orthebufferofyourchoicewith0.04%Pluronic®F-127.Theexactconcentrationoftheindicatorrequiredforcellloadingmustbedeterminedempirically.
      Note:ThenonionicdetergentPluronic®F-127issometimesusedtoincreasetheaqueoussolubilityofCalbryte™520AM,Calbryte™590AMorCalbryte™630AMesters.AvarietyofPluronic®F-127solutionscanbepurchasedfromAATBioquest.
    3. Ifyourcells(suchasCHOcells)containorganicanion-transports,probenecid(1-2mM)maybeaddedtothedyeworkingsolution(finalinwellconcentrationwillbe0.5-1mM)toreduceleakageofthede-esterifiedindicators.ProbenecidmightnotbeneededeveninCHOcellswithCalbryte™520AM
      Note:AvarietyofReadiUse™probenecidincludingwatersolublesodiumsaltandstABIlizedsolutioncanbepurchasedfromAATBioquest.
    4. Addequalvolumeofthedyeworkingsolution(fromStepborc)intoyourcellplate.
    5. Incubatethedye-loadingplateinacellincubatorfor~60minutes,andthenincubatetheplateatroomtemperatureforanother15minutes.
    6. ReplacethedyeworkingsolutionwithHHBSorabufferofyourchoicethatcontainsananiontransporterinhibitor,suchas1mMprobenecid,toremoveexcessprobes.
    7. RunthecalciumtestsatEx/Em=490/525nmforCalbryte™520AM,540/590nmforCalbryte™590AMor610/640nmforCalbryte™630AM.
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

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