Visualizesubcellularstructuresdirectlyandnoninvasivelybyfluorescencemicroscopy.Withthesevectors,youcanstudycytoskeletalandorganellestructureandfunctioninlivingcells,inrealtime,andwithoutchemicalstaining.Youcanmonitorthelocationofaproteinofinterestrelativetoagivensubcellularstructurebylabelingboththeproteinandthestructurewithseparatefluorescentproteins.Ready-madelentiviralparticlesarealsoavailableforthisapplication.
Visualizesubcellularstructuresdirectlyandnoninvasivelybyfluorescencemicroscopy.Withthesevectors,youcanstudycytoskeletalandorganellestructureandfunctioninlivingcells,inrealtime,andwithoutchemicalstaining.Youcanmonitorthelocationofaproteinofinterestrelativetoagivensubcellularstructurebylabelingboththeproteinandthestructurewithseparatefluorescentproteins.Ready-madelentiviralparticlesarealsoavailableforthisapplication.Thesubcellularlocalizationvectorsencodefusionsoffluorescentproteinvariantsandlocalizationsignalsorsubcellularstructuralproteins,whichtargetthefluorescentproteintoaspecificorganelleorsubcellularstructure.Thevectorsareavailableinavarietyoforganelle-andCytoskeleton-targetedcolorvariants.Theyareidealformultiplexlabelingexperiments.
PhotoactivatablemCherryvectors
PAmCherryisaphotoactivatablemutantofmCherrythatisnonfluorescentuntilitisexposedto350–400nmlight.Byselectingwhichcellularregionstoactivate,youcantrackorganellesagainstadarkbackground.PAmCherrysubcellularlocalizationvectorsareavailablethattargetPAmCherrytothecellmembrane,mitochondria,actin,ortubulin.ObservephotoactivatedPAmCherrywiththesamefiltersetsusedforotherredfluorescentproteins,suchasDsRedvariantsandmCherry.
Lenti-XActinDynamicsMonitoringKit
Thiskitisdesignedtomonitorthehighlydynamicbehavioroftheactinfilamentsysteminlivecells.ThekitincludeslentiviralvectorsencodingactinfusionstoDD-AcGFP1(green,destABIlized)andmCherry(red),andtheDD"sstabilizingligandShield1.DD-AcGFP1-Actiniscontinuouslytargetedfordegradationinthecellbytheproteasomesunlessthecellsareculturedinmediumcontainingthestabilizingligand,Shield1.Bycontrast,mCherry-Actin(whichdoesnotcontaintheDD)hasnormalstabilityuponexpressionandisconstitutivelypresentinthecell.
AddingandremovingShield1createsapulse-chase-likesetofconditionswhichallowyoutomonitorpolymerizationofactinmonomersasnewlysynthesizedDD-AcGFP1-ActinthatisstabilizedasShield1(green)isintegratedintotheexisting(red)mCherry-Actinactinfilamentnetwork.
Autophagysensorvector
pAutophagSENSEisamammalianexpressionvectorthatallowsyoutomonitortheprocessofautophagy.pAutophagSENSEencodesafusionofthegreenfluorescentproteinAcGFP1andtheratLC3protein.Incellsnotundergoingautophagy,theAcGFP1-LC3fusionproteinisevenlydistributedinthecytoplasmofatransfectedcell.However,ifacellisundergoingautophagyviatheformationofautophagosomes,theAcGFP1-LC3fusionproteinisincorporatedintoautophagosomes.Thisprocesscanbemonitoredbyfollowingtheredistributionofthegreenfluorescentfusionproteinfromthecytosoltotheformingautophagosomes.Thereadoutisachangefromanevencytosolicdistributionofgreenfluorescenceincellstoapunctatepatternrepresentingtheformingautophagosomes.pAutophagSENSEsavesyoumoneycomparedtootherautophagydetectionsystems,becauseitisvector-based—notaconsumableyouhavetobuyoverandoveragain.
ebiomall.com
>
>
>
>
>
>
>
>
>
>
>
>
NaturalProteinStopsDeadlyHumanBrainCancerInMice
ScientistsfromJohnsHopkinsandfromtheUniversityofMilanhaveeffectivelyproventhattheycaninhibitlethalhumanbraincancersinmiceusingaproteinthatselectivelyinducespositivechangesintheactivityofcellsthatbehavelikecancerstemcells.ThereportispublishedinNature.
Themostcommontypeofbraincancer-glioblastoma-ismarkedbythepresenceofthesestem-cell-likebraincells,which,insteadoftriggeringthereplacementofdamagedcells,formcancertissue.Stemcells,unlikeallothercellsinthebody,arecapableofformingalmostanykindofcellwhentheright"signals"triggertheirdevelopment.
Fortheirtreatmentexperiment,theresearchersreliedonaclassofproteins,bonemorphogenicproteins,thatcauseneuralstem-cell-likeclusterstolosetheirstemcellproperties,whichinturnstopstheirABIlitytodivide.
Firsttheypretreatedhumanglioblastomacellswithbonemorphogenicprotein4(BMP4),theninjectedthesetreatedcellsintomousebrains.Inmiceinjectedwithcellsthatwerenotpretreated,large,invasivecancersgrew.InthemicewithBMP4-treatedcells,nocancersgrewatall.Threetofourmonthsafterinjection,allmicethatgotuntreatedcellsdied,andnearlyallmicewithBMP4-treatedcellswerealive.
Next,thescientistsdeliveredslow-releaseBMP4-containing"beads"directlyintomousebrainswithimplantedglioblastomacells.Micethatgotemptybeadsdevelopedlargemalignanttumorsanddied.MicewithBMP4beadssurvivedmuchlonger,and80percentsurvivedfourmonthsaftercancercellimplants.
"OurideaistotreatpatientswithBMP4orsomethinglikeitrightaftersurgerytoremoveglioblastomainhopesofpreventingtheregrowthofthecancerandimprovingsurvivaltime,"saysAlessandroOlivi,M.D.,directoroftheDivisionofNeurosurgicalOncologyatHopkinsandacontributortothestudy.
OlivisaysclinicalstudiesusingBMP4couldbeginwithinayearand,ifsuccessful,drugtherapiescouldbeavailabletothepublicwithinthreetofouryears.
"ThiswasproofoftheideathatBMPscouldstopglioblastomabydepletingthestem-cell-likepopulationthatfeedsit,"saysHenryBrem,M.D.,chairmanoftheDepartmentofNeurosurgeryatHopkinsandacollaboratorinthestudy."Thisopensexcitingdoorstofutureresearchintotreatmentsandtherapiesforsuchadevastatingdisease."
