Fluorescein-Diacetate-3-6-Diacetoxyfluoran-DataSheet-生命科学试剂-MedChemExpress

Hotline:400-820-3792Inhibitors•ScreeningLibraries•Proteinswww.MedChemEFluoresceinDiacetateCat.No.:HY-D0719CASNo.:596-09-8Synonyms:3,6-Diacetoxyfluoran;Di-O-acetylfluorescein分⼦式:C₂₄H₁₆O₇分⼦量:416.38作⽤靶点:Others作⽤通路:Others储存⽅式:-20°C,protectfromlight*Insolvent:-80°C,6months;-20°C,1month(protectfrom

light)溶解性数据体外实验DMSO:26.67mg/mL(64.05mM;Needultrasonic)MassSolvent1mg5mg10mgConcentration制备储备液1mM2.4017mL12.0083mL24.0165mL5mM0.4803mL2.4017mL4.8033mL10mM0.2402mL1.2008mL2.4017mL请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；⼀旦配成溶液，请分装保存，避免反复冻融造成的产品失效。储备液的保存⽅式和期限：-80°C,6months;-20°C,1month(protectfromlight)。-80°C储存时，请在6个⽉内使⽤，-20°C储存时，请在1个⽉内使⽤。体内实验请根据您的实验动物和给药⽅式选择适当的溶解⽅案。以下溶解⽅案都请先按照InVitro⽅式配制澄的储备液，再依次添加助溶剂：(为保证实验结果的可靠性，澄的储备液可以根据储存条件，适当保存；体内实验的⼯作液，建议您现⽤现配，当天使⽤；以下溶剂前显⽰的百分⽐指该溶剂在您配制终溶液中的体积占⽐；如在配制过程中出现沉淀、析出现象，可以通过加热和/或超声的⽅式助溶)1.请依序添加每种溶剂：10%DMSO>>90%(20%SBE-β-CDinsaline)Solubility:≥2.08mg/mL(5.00mM);Clearsolution1/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemE2.请依序添加每种溶剂：10%DMSO>>90%cornoilSolubility:≥2.08mg/mL(5.00mM);ClearsolutionBIOLOGICALACTIVITY⽣物活性Fluoresceindiacetate⼀种细胞渗透性的酯酶底物。Fluoresceindiacetate可作为⼈⾕胱⽢肽硫转移酶Pi(hGSTP1-1)的荧光底物。体外研究Fluoresceindiacetate(FDA)isanacetylatedderivativeofthegreenfluorescentdyefluorescein[1].Fluoresceindiacetate(FDA),afluorescentprobeusedforvitalstaining,isafluorescentlyactivatedbyesterolyticactivityofhumanPi-classglutathioneS-transferase(hGSTP1)selectivelyamongvariouscytosolicGSTs.FluorescenceactivationofFluoresceindiacetatesusceptibletoGSTinhibitorsisobservedinMCF7cellsexogenouslyoverexpressinghGSTP1,butnotincellsoverexpressinghGSTA1orhGSTM1.FluoresceindiacetatecanbeusedasafluorogenicsubstrateforhGSTP1-1.ToinvestigatewhetherthefluorescenceactivationisduetohGSTP1activity,FluoresceindiacetateisincubatedwithrecombinanthGSTP1-1andGSHinvitro.RemarkablefluorescenceactivationisobservedinthepresenceofbothhGSTP1-1andGSH,whereasonlyslightactivationisobservedintheabsenceofeitherofthemorwhentheenzymeisheatinactivated.ThissuggeststhatthefluorescenceactivationofFluoresceindiacetatedependsonhGSTP1-1activity.FromthelinearrelationshipbetweentherateofincreaseinfluorescenceandthehGSTP1-1concentration,thespecificactivityoftheenzymefor1μMFluoresceindiacetateisdeterminedtobe79±15nmol/min/mgprotein.FluoresceindiacetateisapplicableasafluorogenicsubstrateforevaluatinginhibitorsofGSTP1-1invitro.ForFluoresceindiacetateasasubstrate,bothEthacrynicacid(EA)andNBDHEXsuppressthehGSTP1-1-dependentfluorescentincreaseinaconcentration-dependentmanner,withIC50sof3.3±0.5μMand0.61±0.04μM,respectively[2].PROTOCOLCellAssay[2]MCF7cells(2-3×105)areseededina35-mmglassbottomdishbeforetheexperiment.Priortoimaging,cellsarewashedwith1mLPBS,thenincubatedin1mLHanks’BalancedSaltsolution(HBSS(+)withoutphenolred)containing1μMFluoresceindiacetate(0.1%DMSOasacosolvent)for5minat37°C.Cellsarewashedtwicewith1mLPBSand1mLHBSSisaddedbeforeimaging.FluorescenceimagesoffluoresceinandDsRed-Express2wereacquiredintheFITCchannel(excitationat473nm)andtheDsRedchannel(excitation559nm).The16-bitimagesobtainedareanalysed[2].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.户使⽤本产品发表的科研⽂献•CellDeathDiscov.2021Dec4;7(1):374.Seemorecustomervalidationsonwww.MedChemE2/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemEMcePdfHeightCaution:Prod