Can anyone suggest a descent method for the determination of k_off for GPCR - small molecule complex that doesn't involve radioactive labeling of the ligand?
Surface plasmone resonance (Biacore) - if your ligand can be coupled to any of the available chip-surfaces and if you want to test few ligands (I assume it is hard to use this for a screening approach).
I've just looked up SPR - GPCR combination in literature and it seems to me that it would take more efforts than radiosynthesis.
Seems like I'll stick with reference radioligand competitive binding assays.
Dear Viachaslau, you can use Microscale Thermophoresis (MST) it is a label-free, free-solution and immobilization-free method: http://onlinelibrary.wiley.com/doi/10.1002/anie.201204268/abstract
Compared with SPR the handling of MST is straight forward and you can, if needed, use your native proteins without any modifications.
Thank you for the suggestion. I'm always happy to see new methods. However, the utility of MST for kinetic measurements is still to be validated, as far as I have understood from your paper.
- Badges
- Science topic
- Similar topics
- Chemistry
- Pharmacology