I have been working in electrochemical immunosensors. I have tried to immobilize anti human IgG from rabbit from sigma aldrich (containing 0.1% sodium azide as preservative) on an electrode surface using EDC NHS coupling. But it didnot bind. I would like to know whether sodium azide will inhibit antibody conjugation.
Although sodium azide is a very potent Ab preservative, it may inhibit EDC/NHS coupling towards antibody. Usually, for the EDC coupling, pH, buffer, temperature, and reaction time will be the main parameter to earn the successful coupling efficiency. Any reactive (charged) molecules such as sodium azide would be better to be removed in advance. You may a take separation/purification step to remove it by dialysis.
Also, the coating efficiency by EDC coupling in a solid surface (electrode) have been discussed earlier, however it is still not that reliable and robust.
Sodium azide can have numerous effects to binding. I am not so sure about EDC/NHS but it definitely affects thiol chemistry. Its worth a try to see its effect on EDC/NHS. Probably trying higher pH can act positive to EDC/NHS coupling
- Badges
- Science topic
- Similar topics
- Biological Science
- Immunology
- Antibodies