| Overview |  PrinterFriendlyVersion |
| Ex/Em(nm) | 485/590 |
| MW | N/A |
| CAS# | N/A |
| Solvent | N/A |
| Storage | R/D/L |
| Category | ProteinBiochemistry Generalproteins |
| Related |
- PrepareProlite™OrangeWorkingSolution:Add1µLof200XProlite™Orange(ComponentA)to199µLofSampleDilutionBuffer(ComponentE)foreachsampleorBSAstandarddetection.
Note:Donotmixtheworkingsolutioninaglasscontainer. - Runproteinassay:
- Add190μL/wellofProlite™OrangeWorkingSolution(fromStep1)intoeachtube.
Note:Usethin-wall,polypropylene,clear0.5mLPCRtubessuchasInvitrogen™Qubit®AssayTubes(Cat#Q32856)orAxygenPCR-05-Ctubes(VWR,Cat#10011-830).Othertypesoftubescanhaveautofluorescenceandmayinterferewiththeassay. - Add10µLBSAstandards(ComponentB,C,D)or10µLsamplesintothe190μLProlite™OrangeWorkingSolutiontube(fromStep2.1)tomakethefinalassayvolume200μL/tube.
- Incubatethereactionatroomtemperaturefor15minutes.Protectthesamplesfromlightandavoidholdingthesamplesinhands.
- InsertthesamplesintoQubit®andmonitorthefluorescencefollowingQubit®manufacture’sinstructionsasbrieflysummarizedbelow.
a)PressProteinontheHomescreenoftheQubit®HomescreenandproceedtopressReadstandards.
b)Inserteachofthe3tubescontainsstandardsintothesamplechamber.
c)ClosethelidandpressReadstandards.
d)Theinstrumentdisplaystheresultsandgeneratescalibrationcurve.
e)PressRunsamplesandselectsamplevolumeto10µL.
f)Insertthesampletubeintothesamplechamber.
g)ClosethelidandpressReadtube.
h)Theinstrumentdisplaystheresultsontheassayscreen.Thetopvalueistheoriginalsampleconcentrationand
bottomvalueisthedilutedconcentration.
Note:FormoredetailedoperatinginformationonQubit®,refertotheQubit®FluorometerUserGuide. - Add190μL/wellofProlite™OrangeWorkingSolution(fromStep1)intoeachtube.
| References&Citations |  PrinterFriendlyVersion |
1. Xu,D.D.;Liu,C.;Li,C.Y.;Song,C.Y.;Kang,Y.F.;Qi,C.B.;Lin,Y.;Pang,D.W.;Tang,H.W.,DualAmplificationFluorescenceAssayforAlphaFetalProteinUtilizingImmunohybridizationChainReactionandMetal-EnhancedFluorescenceofCarbonNanodots.ACSApplMaterInterfaces2017,9(43),37606-37614.
2. Li,J.;Qiu,X.J.,QuantificationofMembraneProteinSelf-AssociationwithaHigh-ThroughputCompatIBLeFluorescenceAssay.Biochemistry2017,56(14),1951-1954.
3. Samokhvalov,A.V.;Safenkova,I.V.;Eremin,S.A.;Zherdev,A.V.;Dzantiev,B.B.,UseofanchorproteinmodulesinfluorescencepolarisationaptamerassayforochratoxinAdetermination.AnalChimActa2017,962,80-87.
4. Akbar,S.M.;Sreeramulu,K.;Sharma,H.C.,Tryptophanfluorescencequenchingasabindingassaytomonitorproteinconformationchangesinthemembraneofintactmitochondria.JBioenergBiomembr2016,48(3),241-7.
5. Jang,E.;Kim,M.;Koh,W.G.,Ag@SiO2-entrappedhydrogelmicroarray:anewplatformforametalenhancedfluorescence-basedproteinassay.Analyst2015,140(10),3375-83.
6. Song,W.;Wang,Y.;Liang,R.P.;Zhang,L.;Qiu,J.D.,Label-freefluorescenceassayforproteinkinasebasedonpeptidebiomineralizedgoldnanoclustersassignalsensingprobe.BiosensBioelectron2015,64,234-40.
7. Caers,J.;Peymen,K.;Suetens,N.;Temmerman,L.;Janssen,T.;Schoofs,L.;Beets,I.,CharacterizationofGprotein-coupledreceptorsbyafluorescence-basedcalciummobilizationassay.JVisExp2014,(89),e51516.
8. Krishna,S.N.;Luan,C.H.;Mishra,R.K.;Xu,L.;Scheidt,K.A.;Anderson,W.F.;Bergan,R.C.,Afluorescence-basedthermalshiftassayidentifiesinhibitorsofmitogenactivatedproteinkinasekinase4.PLoSOne2013,8(12),e81504.
9. Zhang,C.;Zheng,L.;Nurnberg,J.;Vacari,B.M.;Zhou,J.;Wang,Y.,Cleavageofpro-tumornecrosisfactoralphabyADAMmetallopeptidasedomain17:afluorescence-basedproteaseassaycleavesitsnaturalproteinsubstrate.AnalBiochem2014,445,14-9.
10. Veiksina,S.;Kopanchuk,S.;Rinken,A.,Buddedbaculovirusesasatoolforahomogeneousfluorescenceanisotropy-basedassayofligandbindingtoGprotein-coupledreceptors:thecaseofmelanocortin4receptors.BiochimBiophysActa2014,1838(1PtB),372-81.
