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Altogen/Astrocyte Transfection Reagent/1.5 ml CRISPR(Catalog #2110)

Description

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TransfectionReagentforAstrocytes(PrimaryAstrocyte Cells,Astroglia)

  • Proprietarycationiclipidsformulation
  • HightransfectionefficiencyofsmallRNA(siRNA,shRNA,miRNA),mRNA,pDNA
  • Effectiveandrobustintracellulardelivery
  • KitincludesTransfectionEnhancerreagent
  • Producesconsistentresults,lot-to-lot,plate-to-plate,andwell-to-well
  • Work inthepresenceofserum
  • Aprovenreagentforestablishingstablecelllines
  • Optimizedtransfectionprotocolsareadaptedforusewithbothstandard&reversetransfectionmethods
  • DownloadinvitroAstrocytetransfectionprotocol:[PDF]
  • Download AstrocyteCRISPR/Cas9transfectionprotocol:[PDF]
  • DownloadPowerPointpresentationforAstrocytecellstransfectionkit:[PPT]
  • DevelopedandmanufacturedbyAltogenBiosystems

TransfectionEfficiency:

Reagentexhibitsatleast70%transfectionefficiencyofsiRNAdelivery.TransfectionefficiencywasdeterminedbyqRT-PCR.

TransfectionProtocolandMSDS:

DownloadAltogenBiosystemsAstrocyteTransfectionProtocol:[PDF]

DownloadMSDS:[PDF]

AstrocyteCells(PrimaryAstrocytesandAstroglia):

Thehumanbraincomprisestwomajorcelltypes:neuronsandglia.NeuronsareresponsIBLefortransmittingnervesignalswhilegliasafeguardneurons.Astrocytes,asortofglialcells,makeover50%ofallbraincells.IntheeventofanydistresstotheCentralNervousSystem(CNS),aprocesscalledgliosisistriggered.Gliosisinvolvestherapiddifferentiationofglialcellstoprovidemechanicalsupporttoneurons.However,excessivegliosisissymptomaticofvariousformsofbraincancer.PerthestudypublishedintheJanuary2016issueofTumorBIOLOGy,astrocytesplayanimportantroleincontrollingtheenvironmentaroundatumor.Astrocytesretardthedetectionofcancercellsbytheimmunesystemandhelpenhancetheirproliferation.Preclinicalresearchtargetingastrocytescanprovidevaluableinsightintomolecularpathwaysthatcancurbtumors.Transfectionisawell-regardedtechniquetoconductpreclinicalresearchinmanydifferentcells,includingastrocytes.

Astrocytesarestar-shapedglialcellsinthebrainandspinalcord.Astrocytesaresupportivecells,andtheirfunctionsincludebiochemicalsupportofendothelialcells,whichformtheblood-brainbarrier,providingnutrientstothenervetissue,extracellularK+levelregulation,neurotransmittersremoval,andhelpingrepairthebrainandspinalcordfollowingtraumaticinjuries.Theyarealsodependentonmyelinationandsynapsesformation.ManyprimaryastrocytesexpresstheintermediatefilamentglialfibrillaryacidicproteinorGFAP.Researchindicatesthatastrocytescommunicatewithneuronsthroughthereleaseofgliotransmittersviaacalciumionprocess.Therearethreetypesofastrocytesinthecentralnervoussystem:fibrous,protoplasmicandrADIal.Itisbelievedastrocytescellbodiesareseparateanddonotoverlap.

Astrocytes,alsoknownasastroglia,arecellsinlivingorganismsthatmakeuppartsofthenervoussystem,particularlyinthebrainandspinalcord.Theyworkwiththebody’snaturalresponsetotraumainthebrainandspinalcord,byrepairingandprovidingnutrientstoaffectedareas.Becauseofthis,astrocytesmaybeusefulinthestudyofspecificmedicaltreatmentsforpatientssufferingfromspinalcordinjuries.Thesecellsarecrucialtoresearchinthefieldofneuroscience.

Primarycellculturesareusedinbiologicalandgenetherapystudiesandserveasvaluablemodelsystemsthatmaymoreaccuratelyrepresentthebiologyofhealthycells.Manyculturedcelllines,aswellasthemajorityofprimarycellcultures,canbetransfectedwithexogenousnucleicacidswhenappropriatetransfectionapproachesareemployed.Sincethemajorityoftransfectionmethodscausessignificanttoxicityinprimarycellcultures,optimizingthisprocedure,specificallytheprotocolandreagentstobeutilized,isessentialfordevelopinganeffectivetransfectionstrategiesforagivencelltype.ALTOGEN®Kitsforprimarycellsandsensitivecelllineshavebeendesignedtohavesignificantlylowercytotoxicitythanotheralternatives.

Data:

Figure1.SiRNAstargetingLaminA/CmRNAornon-silencingcontrolsiRNAweretransfectedintoactrocytecellsfollowingtherecommendedprotocol.At48hourspost-transfectionthecellswereanalyzedbyqRT-PCRforgeneexpressionlevels.18SrRNAlevelswereusedtonormalizetheLaminA/Cdata.Valuesarenormalizedtountreatedsample.Dataaremeans±SD(n=3).

Astrocyte-cells-transfection-protocol

Figure2.ProteinexpressionofLaminA/CinAstrocytecells.DNAplasmidexpressingLaminA/CorsiRNAtargetingLaminA/CweretransfectedintoAstrocytecellsfollowingAltogenBiosystemstransfectionprotocol.At72hourspost-transfectionthecellswereanalyzedbyWesternBlotforproteinexpressionlevels(normalizedbytotalprotein,10µgoftotalproteinloadedpereachwell).Untreatedcellsusedasanegativecontrol.

SelectedAstrocyteTransfectionReagent productcitations:

  • JournalofNeuroscience.2013 33(44).Foxo3aTranscriptionallyUpregulatesAQP4andInduces …Kapooretal [PDF]
  • MolCellBiol.201333(7).SCO2inducesp53-mediatedapoptosisbyThr845phosphorylation…Madanetal[PDF]

RequestaQuotefor
AstrocyteTransfectionReagent

AltogenBiosystems:

AltogenBiosystemsprovidespre-optimizedtransfectionkitsandelectroporationproductsforlifesciencesandcancerresearch.Transfectionprotocolsareoptimizedforindividualcancercelllines.AltogenBiosystemsdevelopedtwotypesofinvivodeliverykits(animaltransfection):1)Tissue-targetedreagents(deliveryofproteins,DNA,andRNAintoliver,pancreas,orkidneytissues),and2)Broadrangeinvivobiodistributionreagents(PEG-Liposomebasedreagent,Nanoparticle-basedinvivoreagent,Lipid-basedtransfectionkit,andPolymer-basedkit).Advancedformulationofreagentsandoptimizedtransfectionprotocolsprovideefficientintracellulardeliveryofproteins,DNA,mRNA,shRNA,siRNA,andothernegativelychargedbiomoleculesinvitroandinvivo.Readmore abouttransfectiontechnologyatAltogen’sTransfectionResource.

AltogenResearchServices:

AltogenLabsprovidesGLP-compliantcontractresearchstudiesforpre-clinicalresearch,INDapplications,anddrugdevelopment.BiologyCROservices include:Xenograftmodels(30+),developmentofstablecelllines,ELISAassaydevelopment,cell-basedandtissuetargetedRNAistudies,safetypharm/toxassays,andotherstudies(visitAltogenLabs.com).

VolumeOptions:

  • 0.5ml(Catalog#1711)
  • 1.5ml(Catalog#1712)
  • 1.5mlCRISPR(Catalog#2110)
  • 8.0ml(Catalog#1713)
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