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主营:研究并生产荧光和发光探针,信号转导研究的试剂
℡ 4000-520-616
℡ 4000-520-616
AAT Bioquest/Portelite™ Fluorimetric Protein Quantitation Kit/11110/200 Tests
产品编号:11110
市  场 价:¥1900.00
场      地:美国(厂家直采)
联系QQ:1570468124
电话号码:4000-520-616
邮      箱: info@ebiomall.com
美  元  价:$95.00
品      牌: AAT Bioquest
公      司:AAT Bioquest
公司分类:
AAT Bioquest/Portelite™ Fluorimetric Protein Quantitation Kit/11110/200 Tests
商品介绍
Overview
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Ex/Em(nm)485/590
MWN/A
CAS#N/A
SolventN/A
StorageR/D/L
CategoryProteinBiochemistry
Generalproteins
Related
Proteinquantificationisnecessaryinproteinpurification,electrophoresis,cellBIOLOGy,molecularbiologyandotherresearchapplications.Biuret,Lowry,BCAandBradfordassaysareroutinelyusedforestimatingproteinconcentration.However,thesecolorimetricassaysarelesssensitive,andrequirelargesamplevolumetoensurehigheraccuracy.OurPortelite™FluorimetricProteinQuantitationKitissignificantlymoresensitivethanexistingstandardcolorimetricmeasurements,e.g.,BradfordandBicinchoninicacid(BCA)assays.Prolite™Orangeusedinthekitisintrinsicallynon-fluorescentinaqueoussolution,butreactsrapidlywithproteinsandgeneratesbrightfluorescence.Portelite™FluorimetricProteinQuantitationKitprovidesarapidmethodforquantifyingproteinconcentrationinsolutions.Aslittleas50ng/mLofBSAcanbedetected.Thekitcanbeperformedwiththequbit®Fluorometer.Itcanbecompletedwithin15minuteswiththefluorescencesignaleasilymonitoredatEx/Em=485/590nm.

Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds.
  1. PrepareProlite™OrangeWorkingSolution:Add1µLof200XProlite™Orange(ComponentA)to199µLofSampleDilutionBuffer(ComponentE)foreachsampleorBSAstandarddetection.
    Note:Donotmixtheworkingsolutioninaglasscontainer.

  2. Runproteinassay:
    1. Add190μL/wellofProlite™OrangeWorkingSolution(fromStep1)intoeachtube.
      Note:Usethin-wall,polypropylene,clear0.5mLPCRtubessuchasInvitrogen™Qubit®AssayTubes(Cat#Q32856)orAxygenPCR-05-Ctubes(VWR,Cat#10011-830).Othertypesoftubescanhaveautofluorescenceandmayinterferewiththeassay.
    2. Add10µLBSAstandards(ComponentB,C,D)or10µLsamplesintothe190μLProlite™OrangeWorkingSolutiontube(fromStep2.1)tomakethefinalassayvolume200μL/tube.
    3. Incubatethereactionatroomtemperaturefor15minutes.Protectthesamplesfromlightandavoidholdingthesamplesinhands.
    4. InsertthesamplesintoQubit®andmonitorthefluorescencefollowingQubit®manufacture’sinstructionsasbrieflysummarizedbelow.

      a)PressProteinontheHomescreenoftheQubit®HomescreenandproceedtopressReadstandards.
      b)Inserteachofthe3tubescontainsstandardsintothesamplechamber.
      c)ClosethelidandpressReadstandards.
      d)Theinstrumentdisplaystheresultsandgeneratescalibrationcurve.
      e)PressRunsamplesandselectsamplevolumeto10µL.
      f)Insertthesampletubeintothesamplechamber.
      g)ClosethelidandpressReadtube.
      h)Theinstrumentdisplaystheresultsontheassayscreen.Thetopvalueistheoriginalsampleconcentrationand
      bottomvalueisthedilutedconcentration.

  3. Note:FormoredetailedoperatinginformationonQubit®,refertotheQubit®FluorometerUserGuide.
References&Citations
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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.


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