Recently, I detected some proteins in THP1 and primary monocytes after LPS (from 100g/ml to 1000g/ml) treatment for 24h, but the results seem unstable. Sometimes I can see dramatically decrease, sometimes a little and sometimes no any change. I don't know what's the problem and do I need cell synchronization before LPS treatment?
If aim is to detect LPS-inducible proteins, one does not need cell synchronization. It may be helpful to use different time course (0, 30 min, 1 h, 3 h, and 6 h). This is because LPS-treatment is likely to stimulate NF-kB and IRF-inducible genes. These genes are induced in a short period of time (0 to 6 h).
Dear Divaker
My target protein is Bcl-2 and I found two times of significant decrease of Bcl-2 after high concentration of LPS (1ug/ml) for 24h, but now I can not repeat it without changing anything.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Cell Biology
- Culture Cells
- Cell Line