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TransfectionReagentfor A549Cells(LungCarcinomaCells,CCL-185)
TwocomponentformulationenhanceslipidmediatedtransfectionefficiencyOptimizedeasy-to-usetransfectionprotocolprovidedfortransfectionofsiRNA,DNA,mRNA,andmicroRNAKitincludesTransfectionEnhancerreagentandrecommendedtransfectionprotocolHightransfectionefficacyinthe presenceofserumExpandyourRNAiapplicationwithareagentoptimizedfordeliveryofbothsiRNAandplasmidReproducIBLetransfectionresultsWorkswellforstandardreversetransfectionandhigh-throughputapplicationsDownloADInvitroA549transfectionprotocol:[PDF]Download A549CRISPR/Cas9transfectionprotocol:[PDF]DownloadPowerPointpresentationforA549cellstransfectionkit:[PPT]DevelopedandmanufacturedbyAltogenBiosystemsTransfectionEfficiency:
Reagentexhibitsatleast80%transfectionefficiencyofsiRNAdelivery.Transfectionefficiencywasdeterminedbyreal-timeRT-PCR.
TransfectionProtocolandMSDS:
DownloadAltogenBiosystemsTransfectionProtocol:[PDF]
DownloadMSDS:[PDF]
A549CellLine:
LungcancerisconsideredtheleADIngcauseofcancerdeathworldwide,despiteasteadydeclineinthemortalityrateresultingfromwidespreadpublichealthawarenesscampaigns.Newadvancesinlungcancertreatmentapproaches,aswellasimmunotherapyandtargetedtherapies,areessentialintreatingpatientswithadvancedstagedisease.AccordingtoDr.JohnHeymach,chairmanofThoracicOncologyatMDAnderson,approximately160,000peoplediefromlungcancereveryyear,anumbermoresignificantthanthenextthreeprominentcancers(colorectal,breast,prostate)combined.TheA549lungcancercelllinehasbeenusedtoinvestigatetreatmentsforvariousformsoflungcancer,inadditiontootherrespiratoryillnessessuchasasthma.TheA549celllinewasderivedin1972fromthecarcinomatouslungtissueofa58-year-oldCaucasianmale.A549cellsareadenocarcinomichumanalveolarbasalepithelialcellsthataresquamousandresponsibleforthediffusionofsubstancessuchaswaterandelectrolytes,acrossthealveolioflungs.Thiscelllineahypotriploidhumancelllinewiththemodalchromosomenumberof66,occurringin24percentofcells.Therewerealsofrequentinstancesofcellswith64,65,and67chromosomecounts.A549cellshavebeenshowntosynthesizelecithinwithahighpercentageofdesaturatedfattyacidsviathecytidinediphosphocholinepathway.ThesecellsareusedinvitromodelforatypeIIpulmonaryepithelialcellmodelfordrugmetabolism.ImmunoperoxidasestainingshowsthatA549cellsarepositiveforkeratin.Thiscelllineistypicallyusedtoinvestigatenumerousrespiratoryailments,suchasviralinfectionsthatcancauseasthma,tissuedamagelinkedtoasbestosexposure,andsmoking-relatedemphysema.A549cellsgrowasamonolayerandarecapableofsynthesizinglecithin.
Data:
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Figure1.SiRNAstargetingLaminA/CmRNAornon-silencingcontrolsiRNAweretransfectedfollowingtherecommendedprotocol.At48hourspost-transfectiontheA549cellswereanalyzedbyqRT-PCRforgeneexpressionlevels.18SrRNAlevelswereusedtonormalizetheLaminA/Cdata.Valuesarenormalizedtountreatedsample.Dataaremeans±SD(n=3).
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Figure2.ProteinexpressionofLaminA/CinA549cells.DNAplasmidexpressingLaminA/CorsiRNAtargetingLaminA/CweretransfectedintoA549cellsfollowingAltogenBiosystemstransfectionprotocol.At72hourspost-transfectionthecellswereanalyzedbyWesternBlotforproteinexpressionlevels(normalizedbytotalprotein,10µgoftotalproteinloadedpereachwell).Untreatedcellsusedasanegativecontrol.
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SelectedcitationsforA549transfectionreagentusedinthefollowingpublications:
JournalofGeneralVirology.2008 89:1978.Roleofretinoicacidinduciblegene-Iinhuman Liaoetal[PDF]PLoSOne. 2012 (7).PharmacologicalInhibitionofMicrosomalProstaglandin ESynthase-1Finettietal[PDF]MolCellBiol.201333(7).SCO2inducesp53-mediatedapoptosisbyThr845phosphorylationMadanetal[PDF]RequestaQuoteforAltogenBiosystems:
AltogenBiosystemsprovidescelltypespecificandpreoptimizedtransfectionproducts, elecroporationkits,andtargetedinvivodeliveryreagents.Advancedformulationofreagentsandoptimizedtransfectionprotocolsprovideefficientintracellulardeliveryofprotein,DNA,mRNA,shRNA,siRNAandotherbiomolecules.Readmore abouttransfectiontechnologyatAltogensTransfectionResource.
AltogenResearchServices:
AltogenLabsprovidesGLP-compliantcontractresearchstudiesforpre-clinicalresearch,INDapplications,anddrugdevelopment.BIOLOGyCROservices include:Xenograftmodels(30+),developmentofstablecelllines,ELISAassaydevelopment,cell-basedandtissuetargetedRNAistudies,safetypharm/toxassays,andotherstudies(visitAltogenLabs.com).
VolumeOptions:
0.5ml(Catalog#1705)1.5ml(Catalog#1706)1.5mlCRISPR(Catalog#2104)8.0ml(Catalog#7021)