Description

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TransfectionReagentforHCN-1ACells(BrainNeuronalCells,CRL-10442)

• Twocomponentformulationenhanceslipidmediatedtransfectionefficiency
• Optimizedeasy-to-usetransfectionprotocolprovidedfortransfectionofsiRNA,DNA,mRNA,andmicroRNA
• KitincludesTransfectionEnhancerreagentandrecommendedtransfectionprotocol
• Hightransfectionefficacyinthe presenceofserum
• ExpandyourRNAiapplicationwithareagentoptimizedfordeliveryofbothsiRNAandplasmid
• ReproducIBLetransfectionresults
• DownloadHCN-1Atransfectionprotocol:[PDF]
• Download HCN1ACRISPR/Cas9transfectionprotocol:[PDF]
• DownloadPowerPointpresentationforHCN-1Acellstransfectionkit:[PPT]
• DevelopedandmanufacturedbyAltogenBiosystems

TransfectionEfficiency:

Reagentexhibitsatleast76%transfectionefficiencyofsiRNAdelivery.TransfectionefficiencywasdeterminedbyqRT-PCR.

TransfectionProtocolandMSDS:

DownloadAltogenBiosystemsHCN1ATransfectionProtocol:[PDF]

DownloadMSDS:[PDF]

HCN-1ACellLine:

AnumberofstudieshavedemonstratedthesuccessfulapplicationofRNAiinprimarycellculturesandnon-neuronalcelllines;however,theuseofgenesilencinginneuronalcelltypeshasbeennotoriouslydifficultbecauseofseveraltechnicalandBIOLOGicallimitations.NeuronsarethemostresistanttoRNAianddifficulttocharacterizeonthemolecularlevelduetotheirheterogeneousnatureandlimitedLifespan.Methodstoconsiderwhenperformingnucleicaciddeliveryexperimentswithneuronalculturesincludeelectroporation,microinjection,viral-basedmethods,andcertainchemicaltransfectionreagents.AlthoughphysicaldeliverymethodsappeartobeefficientforsiRNA-inducedstudieswheretransientmodificationofgeneexpressionissufficient,thesemethodsdonotseemtobeadequateforbehavioralstudieswherelong-termalterationofgeneexpressionmaybenecessary.Theviral-basedsiRNAexpressionsystemsappeartobemostapplicabletobothCentralNervousSystem(CNS)genetherapyandbasicneurobiologicalstudies.LentiviralvectorshavebeenreportedlyusedsuccessfullyforthedeliveryofshorthairpinRNA(shRNA),aprecursorofsiRNA,intoprimaryneuronstoinduceRNAi.

TheHCN-1Acelllineisahumancorticalneuronallinewasderivedfroman18month-oldfemalewithintractableseizures.Treatmentincludedahemispherectomy.TheHCN-1Acelllinehastheabilitytodifferentiate,displayingmaturemorphologyafterdifferentiationwithsignificantlyslowergrowthrates. HCN-1Aarefrequentlyusedtomodelneuronalprocessesforscientificpurposes. HCN-1Acellshavebeenshowntotestpositiveforsomatostatin,glutamate,neuron-specificenolase,tubulin,vimentin,gamma-aminobutyricacid,vasoactiveintestinalpeptide,andcholecystokinin-8.Thecellstestnegativeforglialfibrillaryacidicprotein,andmyelinbasicprotein.DifferentiationofHCN-1Acanbeinducedthroughculturingthecorticalneuronalcellswithamixtureofnervegrowthfactor,dibutyrylcyclicadenosinemonophosphate,and1-isobutyl-3-methylxanthine.WhenHCN-1Acellsdifferentiate,theytakeonmaturemorphologywhichsignificantlyslowstheirgrowthrate,withadoublingtimeexceeding120hours.AfterHIV/N1Tinfection,HCN-1Adiminishedinsizeandsmallneuriteswereobserved.TransfectionstudiescarriedoutusingtheHCN-1Aneuronalcelllineareanessentialtoolinneurologicalresearch.AltogenBiosystemsprovideshighlyefficientlipid-basedtransfectionreagentkitsfortheHCN-1Acellline.

Data:

Figure1.GAPDmRNAlevelswerequantifiedusingreal-timeRT-PCRintheHCN1AcellstransfectedwithsiRNAstargetingGAPDornon-silencingsiRNA.Forty-eighthourspost-transfection,thecellswereharvestedandanalyzedbyreal-timeRT-PCRforGAPDmRNAexpressionlevels.Datawerenormalizedagainstthe18SrRNAsignal.Controlsampleswereeithermock-transfectedoruntreated.Valuesarenormalizedtountreatedsample.Dataaremeans±SD(n=5).

Figure2.ProteinexpressionofGAPDHinHCN-1Acells.DNAplasmidexpressingGAPDHorsiRNAtargetingGAPDHweretransfectedintoHCN-1AcellsfollowingAltogenBiosystemstransfectionprotocol.At72hourspost-transfectionthecellswereanalyzedbyWesternBlotforproteinexpressionlevels(normalizedbytotalprotein,10µgoftotalproteinloadedpereachwell).Untreatedcellsusedasanegativecontrol.

SelectedHCN1Atransfectionkitcitations:

• JournalofBiologicalChemistry.2011.RegulatoryFactorX1-inducedDown-regulationof Transforming…Fengetal[PDF]
• CancerResearch.201171(15):5144-53.InhibitionofmiR-193aexpressionby…Iliopoulosetal[PDF]
• RNA.201016(11):2108-19.RNaseLreleasesasmallRNAfromHCVRNAthatrefolds…Malathietal[PDF]

AltogenBiosystems:
AltogenBiosystemstransfectionandelectroporationproductsforlifesciencesandcancerresearch.Companydevelopedtwotypesofinvivodeliverykitsforanimalresearch:Tissue-targetedreagents(deliverytoliver,pancreas,andkidneytissues),andbroadrangeinvivodeliveryreagents(PEG-Liposome,Nanoparticle-based,Lipid-based,andPolymer-basedkits). Advancedformulationofreagentsandoptimizedtransfectionprotocolsprovideefficientintracellulardeliveryofproteins,DNA,mRNA,shRNA,siRNA,andothernegativelychargedmoleculesinvitroandinvivo.Readmore abouttransfectiontechnologyatAltogen’sTransfectionResource.

AltogenLabsResearchServices:

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

VolumeOptions:

• 0.5ml(Catalog#1759)
• 1.5ml(Catalog#1760)
• 1.5mlCRISPR(Catalog#2146)
• 8.0ml(Catalog#7043)

Altogen Biosystems是一家开发和制造用于生命科学研究，药物发现和开发的转染试剂盒的生物技术公司。转染试剂盒针对特定癌细胞系和原代细胞培养进行了优化，可将生物分子有效递送到靶组织中。通过先进的试剂配方和优化的转染方案实现体外（癌细胞系）和体内（动物组织靶向试剂、癌细胞系）递送货物分子，包括质粒DNA，各种类型的RNA（mRNA，siRNA，shRNA，microRNA），蛋白质和小分子研究。

苏州蚂蚁淘代理Altogen Biosystems在中国的业务。Altogen Biosystems北京代理，Altogen Biosystems华东代理，Altogen Biosystems华北代理，Altogen Biosystems全国代理，Altogen Biosystems华南代理。