Principle of BacMam Reagents BacMam 2.0 GFP particles enter the cell by endocytosis. After endosomal escape the BacMam DNA moves to the nucleus where only the transgene is expressed; baculovirus genes are not recognized in mammalian cells—hence no virus replication and no toxicity. Transgene expression begins 4-6 hours after transduction. Peak gene expression is generally reached after 24 to 48 h and lasts from 4 to 5 days in transformed cells to more than 2 weeks in slowly dividing cell types. CellLight® Reagent Workflow Using the BacMam 2.0-based CellLight® reagents to express genes is as simple as it is efficient: 1. Add virus to cells in full medium 2. Incubate overnight 3. Image or freeze for future use. Cells can be transduced by mixing cells with the CellLight® reagent immediately prior to plating. Alternatively, adherent cells can be transduced by addition of virus to cells already in dish or on plate. Typically optimal transduction is obtained at 70% confluence and with a 20:1 BacMam particle to cell ratio. Live cell imaging with CellLight™ reagents. Cascade Biologics® human aortic smooth muscle cells (HASMC) were transduced with CellLight™ Lysosomes-RFP (Cat. No. C10597), CellLight™ MAP4-GFP (Cat. No. C10599) and Hoechst 33342 (Cat. No. H24192). Imaging was performed on live cells using a DeltaVision® Core microscope and standard DAPI/FITC/TRITC filter sets. | 
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