I am performing a study in which I am observing the interaction between two proteins following a stimulation. Each protein is tagged with half of a GFP moiety and therefore upon interaction would hope to see a GFP signal. I carry out my stimulations at room temperature and take images every hour. The issue that I am facing is that my unstimulated samples (with split GFP constructs) also show a fair amount of background that increases as I leave the cells at RT (not sure if reconstitution is favorable at RT). How do I go about reducing the background noise?
Artem G Evdokimov
1) engineer your split construct
2) provide decoy (nonfluorescent) protein (the B component of the split pair) in excess - it will 'soak up' the spurious association.
0 votes
0 thanks
- Badges
- Science topic
- Similar topics
- Biological Science
- Cell Biology
More Sushanth Kumar's questions See All
Similar questions and discussions