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AAT Bioquest/Cell Navigator™ Fluorimetric Lipid Droplet Assay Kit *Red Fluorescence*/22735/500 Tests
| Overview |  PrinterFriendlyVersion |
| Ex/Em(nm) | 550/640 |
| MW | N/A |
| CAS# | N/A |
| Solvent | N/A |
| Storage | F/D/L |
| Category | CellBIOLOGy LabelingCells |
| Related | FluorescenceImaging |
| Spectrum | AdvancedSpectrumViewer |
- PrepareDroplite™Redstainingsolution:
- WarmDroplite™Red(ComponentA)toroomtemperature.
- PrepareDroplite™Redstainingsolutionbydiluting2µLofDroplite™Red(ComponentA)to1mLofStainingBuffer(ComponentB).
Note1.20µLofDroplite™Red(ComponentA)isenoughforone96-wellplate.AliquotandstoreunusedDroplite™Redat<-20=""ºc.=""protect=""it=""from=""light=""and=""avoid=""repeated=""freeze-thaw="">
Note2.TheoptimalconcentrationoftheDroplite™Redvariesdependingonspecificapplications.ThestainingconditionsmaybemodifiedaccordingtoaparticularcelltypeandthepermeABIlityofthecellsortissuestotheprobe.
- Prepareandstaincells:
- Foradherentcells:
a.Growcellseitherina96-wellblackwall/clearbottomplate(100µL/well/96-well)oroncover-slipsinsideapetridishfilledwiththeappropriateculturemedium.
b.Gentlyaspiratetheculturemedium,andaddequalvolume(suchas100µL/well/96-wellplate)oftheDroplite™Redstainingsolution(fromStep1.2).
c.Incubatethecellsina37°C,5%CO2incubatorfor10~30minutes.
d.RemoveDroplite™Redstainingsolution(Optional,seeNote1below).
e.ReadFluorescenceat550/640nmwithamicroplatereaderorobservethecellsusingafluorescencemicroscopewithaTRITCfilterset. - ForsUSPensioncells:
a.Centrifugethecellsat1000rpmfor5minutestoget1-5×105cellspertube.
b.Resuspendcellsin500μLofDroplite™Redworkingsolution(fromStep1.2).
c.Incubateatroomtemperatureor37ºCfor10to30min,protectedfromlight.
d.CentrifugetoremovetheDroplite™Redworkingsolution,andresuspendcellsin500μLofpre-warmedmediumorbufferofyourchoicetoget1-5×105cellspertube(Optional,seeNote1below).
e.MonitorthefluorescenceincreaseusingfluorescencemicroscopewithaTRITCfiltersetorflowcytometeratFL1channel.
Note1:SinceDroplite™Redhasminimalfluorescenceinaqueousmedia,aspirationofthegrowthmedium(step2.1b)andremovalofDroplite™Redstainingsolution(step2.1d)afterstainingisoptional.
Note2:Stainedcellscanbefixedwith3-4%formaldehyde.Inaddition,prefixedcells(3-4%formaldehydefixation)canbestainedwithDroplite™Redstainingsolution.
- Foradherentcells:
| References&Citations |  PrinterFriendlyVersion |
1. Nevo-Yassaf,I.;Lovelle,M.;Nahmias,Y.;Hirschberg,K.;Sklan,E.H.,LivecellimagingandanalysisoflipiddropletsbiogenesisinHCVinfectedcells.Methods2017.
2. Makino,A.;Hullin-Matsuda,F.;Murate,M.;Abe,M.;Tomishige,N.;Fukuda,M.;Yamashita,S.;Fujimoto,T.;Vidal,H.;Lagarde,M.;Delton,I.;Kobayashi,T.,Acuteaccumulationoffreecholesterolinducesthedegradationofperilipin2andRab18-dependentfusionofERandlipiddropletsinculturedhumanhepatocytes.MolBiolCell2016,27(21),3293-3304.
3. Wang,C.W.,Lipiddroplets,lipophagy,andbeyond.BiochimBiophysActa2016,1861(8PtB),793-805.
4. Takahashi,H.;Kutasy,B.;Friedmacher,F.;Takahashi,T.;Puri,P.,Expressionofhepaticlipiddropletsisdecreasedinthenitrofenmodelofcongenitaldiaphragmatichernia.PediatrSurgInt2016,32(2),155-60.
5. Schneider,M.R.,Lipiddropletsandassociatedproteinsinsebocytes.ExpCellRes2016,340(2),205-8.
6. Natarajan,S.K.;Rasineni,K.;Ganesan,M.;Feng,D.;McVicker,B.L.;McNiven,M.A.;Osna,N.A.;Mott,J.L.;Casey,C.A.;Kharbanda,K.K.,Structure,functionandmetabolismofhepaticandadiposetissuelipiddroplets:implicationsinalcoholicliverdisease.CurrMolPharmacol2015.
7. Papadopoulos,C.;Orso,G.;Mancuso,G.;Herholz,M.;Gumeni,S.;Tadepalle,N.;Jungst,C.;Tzschichholz,A.;Schauss,A.;Honing,S.;Trifunovic,A.;Daga,A.;Rugarli,E.I.,Spastinbindstolipiddropletsandaffectslipidmetabolism.PLoSGenet2015,11(4),e1005149.
8. Hashemi,H.F.;Goodman,J.M.,Thelifecycleoflipiddroplets.CurrOpinCellBiol2015,33,119-24.
9. Kochan,K.;Maslak,E.;Krafft,C.;Kostogrys,R.;Chlopicki,S.;Baranska,M.,Ramanspectroscopyanalysisoflipiddropletscontent,distributionandsaturationlevelinNon-AlcoholicFattyLiverDiseaseinmice.JBiophotonics2015,8(7),597-609.
10. Billecke,N.;Rago,G.;Bosma,M.;Eijkel,G.;Gemmink,A.;Leproux,P.;Huss,G.;Schrauwen,P.;Hesselink,M.K.;Bonn,M.;Parekh,S.H.,ChemicalimagingoflipiddropletsinmuscletissuesusinghyperspectralcoherentRamanmicroscopy.HistochemCellBiol2014,141(3),263-73.
