optimized-protocol-、cells-without(1).pdf

Journal of Visualized ECopyright 2011 Journal of Visualized ExperimentsPage 1 of 1Materials List for:Optimized Protocol for Efficient Transfection of Dendritic Cells without CellMaturationRobert Bowles*1,Sonali Patil*1,Hanna Pincas1,Stuart C.Sealfon11Center for Translational Systems Biology and Department of Neurology,Mount Sinai School of Medicine*These authors contributed equallyCorrespondence to:Stuart C.Sealfon at stuart.sealfonmssm.eduURL:http:/ NumberCommentsAmaxa Nucleofector 96-wellShuttleLonza Inc.108S0109Serial numberAmaxa Human Monocyte 96-wellNucleofector KitLonza Inc.VHPA-2007Contains the Human Monocyte96-well Nucleofector Solution,the 96-well Supplement and theNucleocuvettes and platesRIG-I siRNADharmaconL-012511-00GLO siRNADharmaconD-001600-01-20RPMI 1640Invitrogen11875Supplemented with 10%FCS,2 mM L-glutamine,100 U/ml penicillin and 100 g/mlstreptomycin to make DC growthmediumDMEMInvitrogen11965L-glutamineInvitrogen25030081Penicillin/streptomycinInvitrogen15070063Fetal Calf SerumHyclone3070.03Dendritic CellsNew York Blood centerDCs are purified from buffy coatsusing a standard procedure