BODIPY-FL-C5-DataSheet-生命科學(xué)試劑-MedChemExpress

Hotline:400-820-3792Inhibitors?ScreeningLibraries?Proteinswww.MedChemEBODIPYFLC5Cat.No.:HY-D1610CASNo.:217075-24-6分?式:C??H??BF?N?O?分?量:320.14作?靶點(diǎn):Others作?通路:Others儲(chǔ)存?式:PleasestoretheproductundertherecommendedconditionsintheCertificateofAnalysis.BIOLOGICALACTIVITY?物活性BODIPYFLC5?種綠?熒光脂酸。BODIPYFLC5可以作為前體合成各種熒光磷脂。BODIPYFLC5對環(huán)境相對不敏感，在?溶性和脂類環(huán)境中都能發(fā)出熒光。體外研究Guidelines(Followingisourrecommendedprotocol.Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds).A.Enzymeassays[1]:1.Analiquotofphospholipidwasplacedina1×1-cmfluorescencecuvetteandbuffer(50mMTris,pH8,100mMNaCl,1mMCaCl2)wasaddedtoatotalvolumeof1.3mL.Thetemperaturewasallowedtoequilibrateat35°Cwithstirringforseveralminutesandthebackgroundemissionwasrecorded(thebackgroundratewasundetectable).Enzyme(1-10μL)wasaddedandtheemissionwasrecorded.2.Analiquotofphospholipid(78μLof0.05mMsubstrate/0.5mMDTPM)wasplacedina1×1-cmfluorescencecuvetteandbuffer(50mMTris,pH8,100mMNaCl,1mMCaCl2,30%glycerol)wasaddedtoatotalvolumeof1.3mL.Thetemperaturewasallowedtoequilibrateat35°Cwithstirringforseveralminutesandthebackgroundemissionwasrecorded(thebackgroundratewasundetectable).Enzyme(1-10μL)wasaddedwithextramixingoftheviscoussolution,andtheemissionwasrecorded.3.AmeasuredamountofBODIPYFL-C5(inthecaseofPBPEC6DNPandMBPEDNP)orBODIPY-FL-C5-lyso-PAF(inthecaseofBC11-DNPC8-PC)wasaddedtothephospholipidsubstratesolutionsandtheincreaseinemissionwasrecorded.Thefactor,picomolesofBODIPYproduct/intensityunitincrease,wasthenusedtoconvertemissionincrease/sectopicomolesofproduct/secondintheassays.B.Zebrafishinvitroassay[1]:1.Embryoswereplacedin0.15mLofembryomedium(EM:13.7mMNaCl,0.537mMKCl,0.025mMNa2HPO4,0.044mMKH2PO4,1.30mMCaCl2,1mMMgSO4,4.2mMNaHCO3,pH7.2).1/2MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemE2.Containing150-200ngofafluorescentphospholipidsubstrate,andsonicatedfor2-5s.3.After1hat37℃,reactionswerestoppedbytheadditionof0.45mLofchloroform:methanol(2:1),mixed,andcentrifuged(30s,16,000g).4.Theaqueousfractionwasdiscardedandanaliquotoforganicfractionwasloadedonthin-layerchromatographyplates.5.Platesweredevelopedintoluene:diethylether:ethanol:aceticacid(50:40:2:0.2)andquantifiedusingalaserscanner.REFERENCES[1].HSHendrickson,etal.IntramolecularlyquenchedBODIPY-labeledphospholipidanalogsinphospholipaseA(2)andplatelet-activatingfactoracetylhydrolaseassaysandinvivofluorescenceimaging.AnalBiochem.1999Dec1;276(1):27-35.McePdfHeightCaution:Producthasnotbeenfullyvalidatedformedicala