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ExpressionSystems’TransfectionMediumisdesignedtocomplementESF921andESFAFinsectcellculturemediafortheco-transfectionortransfectionstepinbaculovirusvectorproduction.TransfectionMediumisaserum-freeandanimal-freeformulationthatfacilitatesvectorDNAuptakebytheinsecthostcells.
ExpressionSystems’TransfectionMediumisdesignedtocomplementESF921andESFAFinsectcellculturemediaforthecotransfectionortransfectionstepinbaculovirusvectorproduction.TransfectionMediumisaserum-freeandanimal-freeformulationthatfacilitatesvectorDNAuptakebytheinsecthostcells.RealizehighertitersbyusingaformulationdesignedtoworkwithESF921andESFAF.
ExpressionSystems’TransfectionMediumwascomparedtooldertechnologythatusesGrace’smediumfortheperformanceofcotransfectiontoproducebaculovirusvectors.SerumFreeMediumwasincludedasacontrol.ResultingvirustitersfromthecotransfectionwereseverallogshigherusingTransfectionMediumascomparedtoothermedia.TransientexpressionwasalsodeterminedasameasureofDNAuptake.Again,TransfectionMediumresultedinsignificantlybetterresultswiththepercentageofgp64expressingcellsbeingatleasttwo-foldhigherthanothermedia.
Sf9cellsweregrowninSerumFreeMediumandplatedintoindividualwellsofadeepwellblockataconcentrationof2x106cellsper100μlperwell.DNAandalipofectionreagentwereincubatedtogetherin200μloftestmedia(eitherSerumFreeMedium,Grace’sMediumorExpressionSystems’TransfectionMedium)for30minutes.DNAmixturewasaddedtocellsandvolumewasbroughtupto1mlintestmedia.CellsUSPensionwasincubatedforfourhoursat27°Cwhileshaking.Attheendoftheincubation,thevolumeofeachwellwasbroughtupto4mlwithSerumFreeMediumandculturedforfourdays.DeterminationoftheP0(i.e.,productofthecotransfection)infectiousunits(IU)viraltiterofthesupernatantwasperformedusingthegp64flowcytometricmethod.Transientexpressionofgp64wasdeterminedfortheculturedcellsbystainingwithgp64-PE.
| Mediatype | Serum-free,animal-freeliquid,1X |
| Shippingcondition | Ambient |
| Storagecondition | 2-8⁰C,protectfromlight |
| Use-bydate | Oneyearfromdateofmanufacture |
SouthernBiotech/Goat Anti-Human Lambda-Alexa Fluor® 555/2070-32/1.0 mg
vectorlabs/Biotinylated Aleuria Aurantia Lectin (AAL)/B-1395/1 mg
vectorlabs/VECTASTAIN® Elite® ABC-HRP Kit (Peroxidase, Mouse IgG)/PK-6102/1 kit
vectorlabs/VECTASTAIN® Elite® ABC-HRP Kit (Peroxidase, Universal)/PK-6200/1 kit
vectorlabs/Unconjugated Musa Paradisiaca (Banana) Lectin (BanLec)/L-1410/5 mg
GiottoBiotech/Calmodulin N60D/5 mg/G02CLM60cn