优点：

数据：

Human osteosarcoma (U2-OS) cells expressing CellLight®Lysosomes-GFP (catalog # C10507) were treated with 200 µM tert-Butyl hydroperoxide for 2 hr. A stain solution containing 5 µM CellROX® Orange (Catalog # C10443) and 2 drops/ml of NucBlue™ Live Cell Stain (Catalog # R37605) was applied for 30 min at 37°C. Cells were washed and imaged with Live Cell Imaging Solution (catalog # A14291DJ).

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.

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

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