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View additional product information for CellLight™ Early Endosomes-RFP, BacMam 2.0 - FAQs (C10587)
9 product FAQs found
与许多其它细胞相比,神经元转导更为困难。提高转导效率的主要方法是采用更多数量的病毒颗粒标记细胞。对于原代神经元,在铺盘时转导要比已培养细胞转导效果更好。此外,蛋白的表达在神经元中发生较慢,表达高峰通常发生于2-3天后而非转导后16小时。
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).
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
Cells transduced with the CellLights reagents can be stored frozen for several months after transduction, without loss of expression.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
If the viral particles are used at the level we recommend, they are very well tolerated by cells.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
The BacMam 2.0 CellLights typically express for 5 days after transduction.
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The first generation BacMam reagents were shown to efficiently transduce over 90 cell types, including stable cell lines and primary cells. With BacMam 2.0, it is now possible to also efficiently transduce primary neurons and stem cells.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
Our BacMam constructs do not replicate in mammalian cells, making them Biosafety Level I (the lowest biosafety level). They can be handled with the standard precautions used for any cell-based reagents.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
BacMam delivery technology works with over 90 mammalian cell types. However, cells with hematopoietic origin do not show effective transduction with CellLight BacMam 2.0 reagents.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
Neurons are more difficult to transduce than many other cells. The main way to improve transduction is to label with a higher number of particles per cell. For primary neurons, it can also help to transduce them at the time of plating rather than on established cultures. There can also be a slower onset of expression in neurons and peak expression often occurs on day 2-3 rather than 16 hours after transduction.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.