ELISA coating question - (Sep/26/2006 )

In capture ELISA, some textbooks mention that coating the primary antibody at 37 degree C for 2 hrs is no different from coating that at 4 degree C overnight. Both process can make the coating successful.

What is the reason behind?
Which one is better?

THX

---

I have never coated or incubated anything at 37C when it comes to ELISA. I have always been told that 2-4 hrs at RT is the same as O/N at 4C. It has to do with binding kinetics of proteins, but I don't know the specifics of how this was determined. I would only assume that 37C would speed up the binding reaction.

---

I would say that when you increase the temperature, you increase the speed and the movements of molecules, increasing the probability to reach the plastic.

---

it just likes the Western Blotting, it only depends on the time when you want to do the experiment. IF u do not have enough time for incubation, then u can incubate at 37 degree C for 2 hours otherwise u can hold it at 4 degree C overnight.

---

it also depends on the stability of the coating protein.

---