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CellLight™ Late Endosomes-RFP, BacMam 2.0 - FAQs

CellLight™ Late Endosomes-RFP, BacMam 2.0 - FAQs

View additional product information for CellLight™ Late Endosomes-RFP, BacMam 2.0 - FAQs (C10589)

9 product FAQs found

CellLight や Premo FUCCI cell cycle sensor などの BacMam 2.0試薬にて、どのようにすれば形質転換効率を上げることができますか？

様々な濃度 (particle-to-cell ratio (PPC))、インキュベーション時の容量、温度、時間、細胞密度を検討してください。 接着細胞の場合、70%程度のコンフレンシーを推奨します。 PPCを調整後、容量が次の重要なパラメーターとなります。 もしそれでもうまくいかない時、BacMam Enhancer Kit (Cat. No. B10107) で効率を上げることもできます。

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.

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.