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View additional product information for CellLight™ FUCCI Cell Cycle Indicator - FAQs (F12345)
8 product FAQs found
Yes. Baculoviruses are insect viruses that do not replicate in mammalian cells and are commonly handled using standard BSL-1 laboratory practices and personal protective equipment appropriate for routine mammalian cell culture work.
Yes. Because BacMam transduction is generally well tolerated, additional BacMam reagent can be added after several days if higher expression levels are needed or if a different BacMam-based assay will be performed. BacMam reagents may also remain continuously present in the culture medium to help maintain expression.
BacMam transduction can often be improved by optimizing the particles per cell (PPC) ratio, incubation volume, incubation time, temperature, and cell density. For adherent cells, we recommend a confluence of 70% or lower at the time of transduction. After PPC optimization, adjusting transduction volume is often the next most effective parameter for improving expression levels. The BacMam Enhancer Kit (Cat. No. B10107) may also help increase expression in some cell types.
Our data show that BacMam-transduced cells can be stored in liquid nitrogen for several months without substantially reducing transgene expression levels after recovery.
BacMam transduction is generally well tolerated, even at high viral particle-to-cell ratios greater than 1,000. However, some BacMam reagents may produce cytotoxic effects at high expression levels depending on the transgene being delivered. High expression of organelle-targeted transgenes may also affect subcellular localization. We recommend optimizing particles per cell (PPC) for each BacMam reagent and cell line combination to balance expression levels and cell health.
Yes. Multiple BacMam reagents can be used simultaneously for co-expression experiments. We recommend pre-mixing the BacMam reagents before adding them to cells and optimizing the particles per cell (PPC) ratio for each reagent combination to achieve balanced expression levels.
BacMam-mediated transgene expression duration depends on factors such as transduction efficiency, cell division rate, mRNA stability, and protein stability. In transformed cell lines such as U-2 OS, HEK293, HeLa, and CHO cells, expression typically lasts about 5 days. In more slowly dividing or contact-inhibited cells, including some stem cells, primary cells, and neuronal cells, expression may persist for more than 2 weeks. In terminally differentiated non-dividing cells, expression has been observed for up to 4 weeks.
BacMam reagents have been shown to efficiently transduce more than 90 cell types, including many stable cell lines and primary cells. Hematopoietic cell lines such as RAW and Jurkat cells generally show lower expression of BacMam-delivered gene products. Because transduction efficiency varies by cell type and target, we recommend optimizing particles per cell (PPC) for each new application to achieve the desired expression level.