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BacMam GFP Transduction Control (BacMam 2.0) - FAQs

查看更多产品信息 BacMam GFP Transduction Control (BacMam 2.0) - FAQs (B10383)

9 个常见问题解答

我使用你们的CellLight BacMam 试剂对我的细胞器进行荧光蛋白标记，但是标记效果不佳。我该如何加以改善？

首先，确定你们的细胞是哺乳动物且非血源性细胞。如果细胞转导效率不佳（例如只有少数细胞被标记或是标记荧光很暗），先试着增加每个细胞的微粒数。接下来试着使用BacMam Enhancer Solution。但是，如果你们的细胞显示标记而标记定位似乎太过普遍或是无处不在，那么你们应该降低每个细胞的微粒数。该手册给出了计算每个细胞的微粒数的指南和公式。

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 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.

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