We have been testing few ELISA kits with human plasma samples, and tried to improve their sensitivity (or signal recovery) by adding to the plasma few concentrations of TRITON X-100 (from 0.4 up to 1%). While some kits did show and improvement, we had problems with a specific one that does seem not to work at all when we add Triton to the samples. Did anyone had the same problems? what can be the reason why that happened?
Thanks!
Detergents like Triton X-100 coat the plastic of the ELISA plate. If you are trying to bind protein directly to the plastic, the detergent will compete for binding sites.
Non-ionic detergent like Triton can suppress non-specific interactions of proteins with each other or with the vessel walls. 1% seems awfully high for this purpose, though, you may get protein unfolding. Try 0.1% instead.
- Badges
- Science method
- Similar topics
- Medicine
- Immunology
- Immunology Techniques
- Immunoassay
- ELISA