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CellLight™ Plasma Membrane-CFP, BacMam 2.0 - FAQs

CellLight™ Plasma Membrane-CFP, BacMam 2.0 - FAQs

查看更多产品信息 CellLight™ Plasma Membrane-CFP, BacMam 2.0 - FAQs (C10606)

10 个常见问题解答

我采用CellLight 标记试剂处理神经元时，转导效率很低，该如何提高转染效率？

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。

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).

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

Is there any way to preserve the CellLights labeling beyond 5 days?

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.

Are the CellLights products toxic to cells?

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.

For how long will the CellLights products label my cells?

The BacMam 2.0 CellLights typically express for 5 days after transduction.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

What cell types can the CellLights products be used with?

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.

What safety precautions need to be taken to work with the BacMam reagent?

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.

Are the products CellLight Golgi-GFP, BacMam 2.0 (Cat. No. C10592) and CellLight Plasma Membrane-CFP, BacMam 2.0 (Cat. No. C10606) classified as S1 or S2 biohazard level?

BacMam is a recombinant version of the AcMNPV baculovirus. It is modified such that it is no longer self-replicating, and thus is classified as BS1 (biosafety level 1) and RG1 (risk group 1 - not known to consistently cause disease in healthy adults, animals or plants, other examples include E. coli and B. subtilis). The resulting BacMam product is a GMO. As such, the "infectious" component of the virus has been removed, thus rendering the agent RG1.

Find additional tips, troubleshooting help, and resources within our Cell Tracing and Tracking Support Center

Do CellLight BacMam 2.0 reagents work on all cell types?

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.

I keep getting low transduction efficiency when using CellLight labeling reagents on my neurons. What can I do to improve the efficiency?

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.