Vivid Colors™ pLenti6.3/V5-GW/EmGFP Expression Control Vector - FAQs

View additional product information for Vivid Colors™ pLenti6.3/V5-GW/EmGFP Expression Control Vector - FAQs (V37006)

4 product FAQs found

I used your pLenti6.3/V5-GW/EmGFP Control Vector and got poor expression of EmGFP after stable transduction into my cell line. What should I do?

This can happen due to excess blasticidin used during selection. Determine the antibiotic sensitivity of your cell line by performing a kill curve. Use the minimum antibiotic concentration required to kill your untransduced cell line.

I used your pLenti6.3/V5-GW/EmGFP Control Vector and got no expression of EmGFP after stable transduction into my cell line. Can you please help?

This could happen due to promoter silencing or using the incorrect filter/detection parameters for flow cytometry. Screen multiple antibiotc-resistant clones and select the one with the highest expression level. Check the filter you are using as well as the FITC detection parameters.

I used your pLenti6.3/V5-GW/EmGFP Control Vector and got poor expression of EmGFP after transient transduction into my cell line. Can you please help?

Please check the following:

- Tranduction occurred in presence of Polybrene reagent
- Use a higher MOI
- Harvest cells at least 48-72 hrs after tranduction

I used your pLenti6.3/V5-GW/EmGFP Control Vector and got no EmGFP-positive cells after titering. What could have happened?

Possible causes include: incorrect filter set/incorrect detection parameters for flow cytometer, viral stocks stored incorrectly, Polybrene reagent not included during transduction, or it may be too soon to see EmGFP expression.