Question (2037)
Question
I transduced abm’s lentivirus into my target cell line but there is no fluorescence or very weak signal. Why?
ABM community
Verified customer
Asked on Mar 24 2025
Answer
There could many possible reasons for this result.
1. Typically GFP is behind a weaker promoter such as SV40 or CBH leading to a less robust fluorescence signal.
2. Virus titer is low leading to only a small proportion of successfully transduced cells. Try increasing the MOI or using a transduction enhancer such as abm’s ViralEntry™ (Cat.No. G516). 3. Target cells are difficult to transduce. A higher MOI must be tested. Some cells are very easy to be transduced such as 293T, while others such as lymphocytes are very difficult.
4. Lentivirus became inactivated during transportation and storage. Lentivirus is very sensitive to temperature changes. Leaving lentivirus at room temperature, 4°C for 48 hours, or subjecting it to multiple freeze/thaws can lead to inactivation or reduced titer.
1. Typically GFP is behind a weaker promoter such as SV40 or CBH leading to a less robust fluorescence signal.
2. Virus titer is low leading to only a small proportion of successfully transduced cells. Try increasing the MOI or using a transduction enhancer such as abm’s ViralEntry™ (Cat.No. G516). 3. Target cells are difficult to transduce. A higher MOI must be tested. Some cells are very easy to be transduced such as 293T, while others such as lymphocytes are very difficult.
4. Lentivirus became inactivated during transportation and storage. Lentivirus is very sensitive to temperature changes. Leaving lentivirus at room temperature, 4°C for 48 hours, or subjecting it to multiple freeze/thaws can lead to inactivation or reduced titer.
ABM Scientific Support
Answered on Mar 24 2025