A time-resolved Förster resonance energy transfer assay to investigate inhibitor binding to ABCG2
This is the biggest paper from my postdoc at the University of Nottingham. We spent years trying to get an FCS assay working on SMALPs containing ABCG2. It showed promise, but kept falling short. In the end, we decided to double-check whether our fluorescent probe was behaving as it should, using a TR-FRET assay. Not only was the probe working, but the double-check assay ended up being surprisingly effective. It turned out in the end that putting ABCG2 in SMALPs interfered with the binding, so the TR-FRET assay became the primary assay we used. Refining the assay took a little while, and we had a real breakthrough about a month from the end of my contract. We also had a really limited amount of the probe available, so the last month of that job was a mad dash of trying to do as many experiments as possible while being as economical with the probe as we could.