4000-520-616
欢迎来到免疫在线!(蚂蚁淘生物旗下平台)  请登录 |  免费注册 |  询价篮
aatbio(优势品牌)
主营:主营:研究并生产荧光和发光探针,信号转导研究的试剂
咨询热线电话
4000-520-616
当前位置: 首页 > 产品中心 > Other_biological_dyes > AAT Bioquest/Cal-520®;,钠盐/21136/1毫克
商品详细AAT Bioquest/Cal-520®;,钠盐/21136/1毫克
AAT Bioquest/Cal-520®;,钠盐/21136/1毫克
AAT Bioquest/Cal-520®;,钠盐/21136/1毫克
商品编号: 21136
品牌: aatbio
市场价: ¥7900.00
美元价: 4740.00
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
PrinterFriendlyVersion

Ex/Em(nm)492/514
MW840.54
CAS#N/A
SolventWater
StorageF/D/L
CategoryCellBIOLOGy
pHandIonIndicators
RelatedBiochemicalAssays
Cal-520®providesarobusthomogeneousfluorescence-basedassaytoolfordetectingintracellularcalciummobilization.Cal-520®AMisanewfluorogeniccalcium-sensitivedyewithasignificantlyimprovedsignaltonoiseratioandintracellularretentioncomparedtotheexistinggreencalciumindicators(suchasFluo-3AMandFluo-4AM).CellsexpressingaGPCRorcalciumchannelofinterestthatsignalsthroughcalciumcanbepreloadedwithCal-520®AMwhichcancrosscellmembrane.Onceinsidethecell,thelipophilicblockinggroupsofCal-520™AMarecleavedbyesterases,resultinginanegativelychargedfluorescentdyethatstaysinsidecells.Itsfluorescenceisgreatlyenhanceduponbindingtocalcium.Whencellsstimulatedwithagonists,thereceptorsignalsthereleaseofintracellularcalcium,whichsignificantlyincreasethefluorescenceofCal-520®.Thecharacteristicsofitslongwavelength,highsensitivity,and>100timesfluorescenceenhancement,makeCal-520®AManidealindicatorforthemeasurementofcellularcalcium.ThehighS/NratioandbetterintracellularretentionmaketheCal-520®calciumassayarobusttoolforevaluatingGPCRandcalciumchanneltargetsaswellasforscreeningtheiragoNISTsandantagonists.Cal-520®sodiumorpotassiumsaltisthehydrolyzedsaltofCal-520®AMincells.Itselectivelybindstocalciumion,andhasthefluorescencethatisstronglydependentoncalciumconcentration.
SpectrumAdvancedSpectrumViewer

Sorry,yourbrowserdoesnotsupportinlineSVG.RelativeIntensity(%)100806040200Sorry,yourbrowserdoesnotsupportinlineSVG.
Sorry,yourbrowserdoesnotsupportinlineSVG.Sorry,yourbrowserdoesnotsupportinlineSVG.
Movemouseovergridtodisplaywavelength&intensityvalues.

300
400
500
600
700
800
900
Wavelength(nm)


Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds.

UseofCal-520®AM,Cal-590™AM,orCal-630™AMEsters

1.LoadCellswithCal-520®,Cal-590™orCal-630™AMEsters:

AMestersarenon-polarestersthatcanreADIlycrosslivecellmembranes,andrapidlyhydrolyzedbycellularesterasesinsidelivecells.AMestersarewidelyusedforloadingavarietyofpolarfluorescentprobesintolivecellsnoninvasively.However,cautionsmustbeexercisedwhenAMestersareusedsincetheyaresusceptibletohydrolysis,particularlyinsolution.Theyshouldbereconstitutedjustbeforeuseinhigh-quality,anhydrousdimethylsulfoxide(DMSO).DMSOstocksolutionsmaybestoreddesiccatedat–20°Candprotectedfromlight.Undertheseconditions,AMestersshouldbestableforseveralmonths.FollowingisourrecommendedprotocolforloadingCal-520®AM,Cal-590™AMorCal-630™AM estersintolivecells.Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds.

a)      Preparea2to5mMstocksolutionofCal-520®AM,Cal-590™AMorCal-630™AMestersinhigh-quality,anhydrousDMSO.

b)      Onthedayoftheexperiment,eitherdissolveCal-520®AM,Cal-590™AMorCal-630™AMinDMSOorthawanaliquotoftheindicatorstocksolutiontoroomtemperature.Prepareadyeworkingsolutionof10to20µMinHanksandHepesbuffer(HHBS)orthebufferofyourchoicewith0.04%Pluronic®F-127.Theexactconcentrationoftheindicatorrequiredforcellloadingmustbedeterminedempirically.

Note:ThenonionicdetergentPluronic®F-127issometimesusedtoincreasetheaqueoussolubilityofCal-520®AM,Cal-590™AMorCal-630™AMesters.AvarietyofPluronic®F-127solutionscanbepurchasedfromAATBioquest.

c)      Ifyourcells(suchasCHOcells)containorganicanion-transports,thenprobenecid(1-2mM)maybeaddedtothedyeworkingsolution(finalinwellconcentrationwillbe0.5-1mM)toreduceleakageofthede-esterifiedindicators.

Note:AvarietyofReadiUse™probenecidincludingwatersolublesodiumsaltandstABIlizedsolutioncanbepurchasedfromAATBioquest

d)      Addequalvolumeofthedyeworkingsolution(fromStepborc)intoyourcellplate.

e)      Incubatethedye-loadingplateinacellincubatorfor60to90minutes,andthenincubatetheplateatroomtemperatureforanother30minutes.

Note:Incubatingthedyelongerthan2hoursgivesbettersignalintensityforsomecelllines.

f)       ReplacethedyeworkingsolutionwithHHBSorifapplicable,abufferofyourchoicethatcontainsananiontransporterinhibitor,suchas1mMprobenecid,toremoveexcessprobes.

g)      RuntheexperimentsatEx/Em=490/525nm(forCal-520®AM),540/590nm(forCal-590™AM)or600/640nm(forCal-630™AM).

2.MeasureIntracellularCalciumResponses:


Figure1.ResponseofendogenousP2YreceptortoATPinCHO-M1cellswithoutprobenecid.CHO-M1cellswereseededovernightat40,000cellsper100µLperwellina96-wellblackwall/clearbottomcostarplate.100µlof4µMFluo-3AM,Fluo-4AMorCal520®AMinHHBSwereaddedintothewells,andthecellswereincubatedat37°Cfor2hour.Thedyeloadingmediumwerereplacedwith100µlHHBS,50µlof300µMATPwereadded,andthenimagedwithafluorescencemicroscope(OlympusIX71)usingFITCchannel.

 

 

  

A                                                                    B

Figure2.ATP-stimulatedcalciumresponseofendogenousP2YreceptorinCHO-K1cellsmeasuredwithCal-520®orFluo-4AM.CHO-K1cellswereseededovernightin50,000cellsper100µLperwellina96-wellblackwall/clearbottomcostarplate.100µLof5µMFluo-4AMortheCal-520®AMwith(A)orwithout(B)2.5mMprobenecidwasaddedintothecells,andthecellswereincubatedat37oCfor2hours. ATP(50µL/well)wasaddedbyFlexStation(MolecularDevices)toachievethefinalindicatedconcentrations.

                            Cal590™AM                                                                                Cal630™AM

    102097-Cell Image Result (High Resolution).bmp     102111-Cell Image Result (High Resolution)-1.bmp          

                        Control                                ATP                                                           Control                            ATP

 

Figure3.ResponseofendogenousP2YreceptortoATPinCHO-Kcells.CHO-Kcellswereseededovernightat40,000cellsper100µLperwellina96-wellblackwall/clearbottomcostarplate.100µlof4µMCal590™AMorCal630™AMinHHBSwith1mMprobenecidwereaddedintothewells,andthecellswereincubatedat37°Cfor2hour.Thedyeloadingmediumswerereplacedwith100µlHHBSand1mMprobenecid,thenimagedwithafluorescencemicroscope(OlympusIX71)usingTRITCchannelbeforeandafteradding50µlof300µMATP.

References&Citations
CitationExplorer
Cal-520®AMhasbeenusedbyresearchersacrosstheglobetoinvestigatetheroleofcalciumionsincriticalbiologicalprocesses.Suchprocessesinclude,butarenotlimitedto,Ca2+channelactivity,Gprotein-coupledreceptorsignalingpathways,intracellular/cytosolicCa2+flux,calciumbursts,actionpotentialsandactivationofcellreceptors.

Below,youmayfindasmallsamplingofspecificCal-520®AMapplications.ToinquireaboutapotentialapplicationofCal-520®AM,ortoconsultwithourfluorescentdyespecialists,pleasecontactusatsupport@aatbio.comor1-800-990-8053.

InNeurobiology,Cal-520®AMhasbeenusedtostudy:
»Neuronsingleactionpotentialsinneocorticalneuronsbothinvitroandinvivo,andassociatedvoltage-dependentcalciumchannels[1]
»Superiorcolliculusinmiceinconjunctionwithtwo-photoncalciumimagingtovisualizeretinalganglioncellsinthesuperficiallamina[2]
»AldolaseCcompartmentsinmice,lookingatcomplexspikesynchronyanditsrelationtosensoryprocessinginawakeanimals[3]
»Neuralcircuitsinbrainsliceandwholebrainpreparation,specificallylookingatindividualactionpotentialsinvivo[4]
»Ca2+dependentcellsignalingpathwaysusingculturedhumanneuroblastomaSH-SY5Ycellsandhigh-speedvideo-microscopy[5]

InCellSignaling,Cal-520®AMhasbeenusedtostudy:
»Intracellularcalciuminspermusingthemicroplatereaderplatformasameanstoquantitatingparameterssuchasmotility[6]
»Calciumsignalingpathwaysinmeniscusfibrochondrocytesbywayofvisualizingcalciumlocalizationandconcentration[7]
»Ca2+mediatedcellularsignalinginrelationtoinositoltriphosphateanditsfluxthroughgapjunctions[8]
»Retinalwave-mediatedformationofcalciumtransientsinMüllerglialcellswithfocusonexpressionofGCaMP3[9]
»Ca2+signalingpathwaysinzebrafishsperm,specificallylookingatcalciumfluxresultingfromcGMP-inducedhyperpolarization[10]

InCardiology,Cal-520®AMhasbeenusedtostudy:
»Sarcoplasmicreticuluminsofarasitsroleincardiacexcitation-contractioncouplingandcalciumsparkevents[11]
»Opticalmappingofcalciumincardiactissueslicestodevelopaframeworkforfutureinvestigationsintocalciumtransients[12]
»Sodium-calciumexchangerfunctionalityandmechanismwithregardstoburstpacemakeractivityinknockoutmice[13]
»Pacemakermodulationinembryonicheartasafunctionofinositol-1,4,5-triphosphatereceptors[14]
»Calciumcurrentandtheroleofpotassiumchannel-interactingprotein2(KChIP2)inmicewithregardstoheartfailure[15]

ViewMoreCitations
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
联络我们
服务热线:4000-520-616
(限工作日9:00-18:00)
QQ :1570468124
手机:18915418616