Originally Posted by msymeonides
Could you explain a bit more, I don't quite understand what the issue was and how you solved it. What is "benchtop time"? Please be as detailed as possible.
The issue was that I have to add samples one by one and this takes 25minutes so the first sample is on the plate for 25minutes before I start the incubation. This 25 minutes difference between adding the first and last sample affected the results. When I extended the imcubation time from 2 till 3 hours I was able to reduce the effect between the first and last sample. The longer the incubation time the closer to equlibrium the ab-ag binding will be the eq and the eq is the same regardless of the start time.
The time the plate is on the benchtop while you prepare it I call bench top time. It is not in the protokoll but you can not avoid having some bench top time. In my case it is normaly 25 minutes when adding samples, as explained earlier. For washing and adding substrates it is only a minute or so.