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。 Altogen Biosystems全国代理部分产品列表： 货号 产品 规格 价格（美元） 5030       1703 A204 Transfection Reagent (Rhabdomyosarcoma) 0.5ml 170 1704 A204 Transfection Reagent (Rhabdomyosarcoma) 1.5ml 348 窗体顶端 窗体底端   2101 A204 Transfection Reagent (Rhabdomyosarcoma) 1.5ml CRISPR 548 窗体顶端 窗体底端   7018 A204 Transfection Reagent (Rhabdomyosarcoma) 8.0ml 1044 窗体顶端 窗体底端   1167 A375 Cells Transfection Reagent (CRL-1619) 0.5ml 195 1168 A375 Cells Transfection Reagent (CRL-1620) 1.5ml 427 2102 A375 Cells Transfection Reagent (CRL-1621) 1.5ml CRISPR 627 1169 A375 Cells Transfection Reagent (CRL-1622) 8.0ml 1281 5030 Nanoparticle In Vivo Transfection Reagent 0.5 ml - 10 injections  445 5010 LIPID In Vivo Transfection Reagent 0.5 ml - 10 injections 405 5040 PEG-Liposome In Vivo Transfection Reagent 0.5 ml - 10 injections 465 5020 POLYMER In Vivo Transfection Reagent 0.5 ml - 10 injections 425 5050 Pancreas In Vivo Transfection Kit 0.5 ml - 10 injections  495 5060 Liver In Vivo Transfection Kit 0.5 ml - 10 injections  495 5070 Kidney In Vivo Transfection Kit 0.5 ml - 10 injections  495 1721 BMS2 Transfection Reagent (Bone Marrow Cells) 0.5 ml  231 6345 C6 Transfection Reagent (Glioma Cells, CCL-107) 0.5 ml  199 6803 DLD-1 Transfection Reagent (Colon Cancer, CCL-221) 0.5 ml  201 1775 HUH-7 Transfection Reagent (Liver Cancer Cells) 0.5 ml  179