How can I increase the transduction efficiency with the BacMam 2.0 reagents such as the the CellLight and Premo products?
Try varying particle-to-cell ratio (PPC), incubation volume, temperature and, cell density (if adherent cells are transduced). For adherent cells, we recommend a confluence of about 70%. Following the PPC, adjusting the volume is the next best parameter to change to optimize protein expression. If that doesn't work, you can also use the BacMam Enhancer Kit (Cat. No. B10107).
I am using your CellLight BacMam reagents for fluorescent protein-labeling of my cell organelles, but they aren't labeling well. How can I improve it?
First, make sure that your cells are mammalian and non-blood derived. If the cells have low transduction efficiency (i.e., only a few cells are labeling or are labeling only dimly), first try increasing the particle number per cell. The next thing to try is to use the BacMam Enhancer Solution. If, however, your cells show labeling, but the localization seems to be too generalized or ubiquitous within the cell, then you should reduce the particle number per cell. The manual gives a guideline and formula for calculating particle number per cell.