Calcein-AM-Calcein-acetoxymethyl-ester-DataSheet-生命科學(xué)試劑-MedChemExpress

Hotline:400-820-3792Inhibitors?ScreeningLibraries?Proteinswww.MedChemECalcein-AMCat.No.:HY-D0041CASNo.:148504-34-1Synonyms:Calceinacetoxymethylester分?式:C??H??N?O??分?量:994.86作?靶點(diǎn):Others作?通路:Others儲(chǔ)存?式:-20°C,protectfromlight*Insolvent:-80°C,6months;-20°C,1month(protectfrom

light)溶解性數(shù)據(jù)體外實(shí)驗(yàn)DMSO:100mg/mL(100.52mM;Needultrasonic)MassSolvent1mg5mg10mgConcentration制備儲(chǔ)備液1mM1.0052mL5.0258mL10.0517mL5mM0.2010mL1.0052mL2.0103mL10mM0.1005mL0.5026mL1.0052mL請(qǐng)根據(jù)產(chǎn)品在不同溶劑中的溶解度選擇合適的溶劑配制儲(chǔ)備液；?旦配成溶液，請(qǐng)分裝保存，避免反復(fù)凍融造成的產(chǎn)品失效。儲(chǔ)備液的保存?式和期限：-80°C,6months;-20°C,1month(protectfromlight)。-80°C儲(chǔ)存時(shí)，請(qǐng)?jiān)?個(gè)?內(nèi)使?，-20°C儲(chǔ)存時(shí)，請(qǐng)?jiān)?個(gè)?內(nèi)使?。BIOLOGICALACTIVITY?物活性Calcein-AM?于測(cè)定細(xì)胞活?的可以滲透細(xì)胞的熒光染料。體外研究Thecalcein-AMdyeusedtostainthelivingcellsisshowntohavealowspontaneousleakageratelessthan15%in4hoursat37°C.Dilutionsoftargetsstainedbycalcein-AMhasalinearrelationshipwithmeasuredfluorescencevalues.NKcells,LAKs,andCTLsarereadilydetectablebythismicrotest.Quantitationofkilling1/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemEandkineticanalysisisreadilyperformedwiththetestsystem[1].Calcein-AMispHindependent,betterretainedandmorephotostable.Inaddition,thehighlevelofintracellularretentionofcalcein-AManditslow-levelreleaseafterincorporationexcludepossiblecell-monolayerlabelingandallowitsuseinacell-cellinteractionassay.Moreover,thebrightfluorescencecaneasilybedetectedandmeasuredbyamicroplatefluorescencereader[2].Calcein-AMisahighlylipophilicvitaldyethatrapidlyentersviablecells,isconvertedbyintracellularesterasestocalceinthatproducesanintensegreen(530-nm)signal,andisretainedbycellswithintactplasmamembrane.Fromdyingordamagedcellswithcompromisedmembraneintegrityorfromcellsexpressingmultidrugresistanceprotein(MRP),unhydrolyzedsubstratesandtheirfluorescentproductsarerapidlyextrudedfromcells.Thecalcein-AMassayhasbeenusedtoassessthecellviability,cytotoxicityandtpquantitateapoptosis[3].體內(nèi)研究Calcein-AMisfoundtobesuitableforinvivostudies,becauseithasnodeleteriouseffectsoncellfunctionandis,indeed,amarkerofcellviability[2].PROTOCOLCellAssay[1][2][3]K562,Daudi,andChanglivercellsarelabeledwithcalcein-AM.Calcein-AM'sexcitationandemissionwavelengthsare496nmand520nm,respectively.Thefilter/mirrorcombinationusedtodetectcalcein-AM'sgreenfluorescenceincludesthe490-nmexcitationand520-nmemissionfilterswithadichroicmirror.Differencesintheautomaticfluorescencereadingsbetweenthetestandcontrolwellsdeterminetheresults[1].Asimpleandsensitivecell-celladhesionmicroplateassayisestablishedusingthecalcein-AM.Theprocedureinvolvesthreesteps:thelabelingoflymphocyteswithanadequateconcentrationofcalcein-AM(20μM)duringashortincubationperiod(30min);theadhesionof2×105labeledlymphocytesperwelltoconfluentkeratinocyteorfibroblastmonolayersgrowninmicrotiterplatesfor90min;and,finally,measurementofthefluorescentsignalutilizinganewsystemofcold-lightmicrofluorimetry[2].Cellsareincubatedfor15minin1mLofa1%saponinsolutioninPBSbuffer,pH7.4,containing0.05%sodiumazide.Aftersaponinpermeabilization,4×105RBCsinsuspensioninPBSbuffercontaining0.1%saponinand0.05%sodiumazideareincubated(37°Cinthedarkfor45min)withcalcein-AMtoafinalconcentrationof5μM,ishedthreetimeswiththesamePBSbuffercontaining0.1%saponinand0.05%sodiumazide,andthecellviabilityisanalyzedbyflowcytometry[3].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.戶(hù)使?本產(chǎn)品發(fā)表的科研?獻(xiàn)?AdvSci(Weinh).2022Oct;9(30):e2203031.?BioactMater.2022Aug11;21:20-31.?ActaPharmSinB.2020Sep;10(9):1694-1708.?RedoxBiol.2020Jan;29:101402.?NanoRes.04May2022.Seemorecustomervalidationsonwww.MedChemEREFERENCES2/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemE[1].WangXM,etal.Anewmicrocellularcytotoxicitytestbasedoncalcein-AMrelease.HumImmunol.1993Aug;37(4):264-70.[2].Braut-BoucherF,etal.Anon-isotopic,highlysensitive,fluorimetric,cell-celladhesionmicroplateassayusingcalceinAM-labeledlymphocytes.JImmunolMethods.1995Jan13;178(1):41-51.[3].BratosinD,etal.Novelfluorescenceassayusingcalcein-AMforthedeterminationofhumanerythrocyteviabilityandaging.CytometryA.20