pTracer™-SV40 Mammalian Expression Vector - FAQs

查看更多产品信息 pTracer™-SV40 Mammalian Expression Vector - FAQs (V87120)

5 个常见问题解答

我使用了你们提供的一款pTracer载体，但未观察到任何荧光。可能是什么原因？

这里列举了一些可能的原因与解决方案：

•培养基中的核黄素导致的高荧光背景——使用1XPBS替换培养基，以减少背景荧光。
•所使用的滤光片组无法使用最佳波长进行激发或无法兼容发射荧光的检测——请使用Omega Optical提供的XF76滤光片。
•转染效率过低以至于无法检测被转染细胞——优化您的转染条件或尝试使用其他方法。
•由于所使用的细胞系无法高表达Cycle 3 GFP蛋白——在HEK-293细胞中，最强的荧光信号通常在转染后72小时出现。

I used one of your pTracer vectors and do not see any fluorescence. What could have happened?

Here are possible causes and solutions:

- High background fluorescence due to riboflavin in the culture medium-replace medium with 1X PBS to eliminate background fluorescence.
- A filter set was used that did not allow excitation at the optimal wavelength or permit detection of the emitted fluorescence-use the XF76 filter from Omega Optical.
- Transfection efficiency is too low to allow detection of transfected cells-optimize your transfection conditions or try another method.
- Expression of Cycle 3 GFP may be low depending on the cell line used-in HEK-293 cells, maximum fluorescence was observed 72 hours post-transfection.

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.

I sequenced one of your vectors after PCR amplification and observed a difference from what is provided online (or in the manual). Should I be concerned?

Our vectors have not been completely sequenced. Your sequence data may differ when compared to what is provided. Known mutations that do not affect the function of the vector are annotated in public databases.

Are your vectors routinely sequenced?

No, our vectors are not routinely sequenced. Quality control and release criteria utilize other methods.

How was the reference sequence for your vectors created?

Sequences provided for our vectors have been compiled from information in sequence databases, published sequences, and other sources.